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Esophageal squamous cell carcinoma (ESCC) is one of the most frequent malignant tumors, and the mechanisms underlying the anti-ferroptosis of esophageal cancer cells are still largely unclear. This study aims to explore the roles of amplified protein kinase C iota (PKCiota) in the ferroptosis of ESCC cells. Cell viability, colony formation, MDA assay, Western blotting, co-IP, PLA, and RNA-seq technologies are used to reveal the roles and mechanisms underlying the PKCiota-induced resistance of ESCC cells to ferroptosis. We showed here that PKCiota was amplified and overexpressed in ESCC and decreased during RSL3-induced ferroptosis of ESCC cells. PKCiota interacted with GPX4 and the deubiquitinase USP14 and improved the protein stability of GPX4 by suppressing the USP14-mediated autophagy–lysosomal degradation pathway. PKCiota was negatively regulated by miR-145-5p, which decreased in esophageal cancer, and also regulated by USP14 and GPX4 by a positive feedback loop. PKCiota silencing and miR-145-5p overexpression suppressed tumor growth of ESCC cells in vivo, respectively; even a combination of silencing PKCiota and RSL3 treatment showed more vital suppressive roles on tumor growth than silencing PKCiota alone. Both PKCiota silencing and miR-145-5p overexpression sensitized ESCC cells to RSL3-induced ferroptosis. These results unveiled that amplified and overexpressed PKCiota induced the resistance of ESCC cells to ferroptosis by suppressing the USP14-mediated autophagic degradation of GPX4. Patients with PKCiota/USP14/GPX4 pathway activation might be sensitive to GPX4-targeted ferroptosis-based therapy. [ABSTRACT FROM AUTHOR]

Moreover, AIF treatment led to cell death via activation of the caspase-associated apoptotic pathway in PCa cells. Cell Growth Analysis To investigate the therapeutic effects of AIF on cell growth, PCa cells were seeded in the 96-well plates (1 × 10 SP 4 sp cells/well) and incubated overnight. As expected, targeting the emerging cancer hallmarks and PCa-specific vulnerability genes, such as FASN and HMGCR [[44]], AIF treatment caused the attenuation of PCa cell growth, migration, and invasion. [Extracted from the article]

Breast tumour kinase (Brk/PTK6) is overexpressed in up to 86% of breast cancers and is associated with poorer patient outcomes. It is considered a potential therapeutic target in breast cancer, even though the full spectrum of its kinase activity is not known. This study investigated the role of the kinase domain in promoting tumour growth and its potential in sensitising triple negative breast cancer cells to standard of care chemotherapy. Triple negative human xenograft models revealed that both kinase-inactive and wild-type Brk promoted xenograft growth. Suppression of Brk activity in cells subsequently co-treated with the chemotherapy agents doxorubicin or paclitaxel resulted in an increased cell sensitivity to these agents. In triple negative breast cancer cell lines, the inhibition of Brk kinase activity augmented the effects of doxorubicin or paclitaxel. High expression of the alternatively spliced isoform, ALT-PTK6, resulted in improved patient outcomes. Our study is the first to show a role for kinase-inactive Brk in human breast tumour xenograft growth; therefore, it is unlikely that kinase inhibition of Brk, in isolation, would halt tumour growth in vivo. Breast cancer cell responses to chemotherapy in vitro were kinase-dependent, indicating that treatment with kinase inhibitors could be a fruitful avenue for combinatorial treatment. Of particular prognostic value is the ratio of ALT-PTK6:Brk expression in predicating patient outcomes. [ABSTRACT FROM AUTHOR]

A near-infrared fluorescent probe, CyAc, is synthesized for accurately diagnosing cancer in vivo by sequential detection of Cys and H+. CyAc can not only achieve a good distinction between normal cells and cancer cells, but also distinguish normal mice from tumor mice. [ABSTRACT FROM AUTHOR]

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Anaesthetics have been implicated to influence cancer cells and progression. Similarly, crosstalk between cancer cells and stromal components within the microenvironment is also an important factor driving progression. Stromal cell-derived factor-1 (SDF-1) and hepatocyte growth factor (HGF) are key chemokines/cytokines produced by fibroblasts which have been established as influential factors in cancer progression. The present study explored the capacity of anaesthetics to influence the expression of these key molecules in fibroblasts. The anaesthetics rocuronium bromide (RB), vecuronium bromide (VB), suxamethonium chloride CRS (SCC), dexmedetomidine hydrochloride (DH) and lidocaine were used to treat MRC-5 fibroblasts over a range of concentrations. Following treatment, transcript expression of SDF-1 and HGF was quantified using quantitative PCR. Treatment of MRC-5 cells with RB brought about a reduction of SDF-1 expression which was found to be significant in the 45 µg/ml treatment group. Treatment with the other anaesthetics brought about some alterations in SDF-1 expression but these were not found to be statistically significant. Treatment with the tested anaesthetics did not have any significant effect on HGF transcript expression within MRC-5 cells, although again some alterations were observed. The results indicated that anaesthetics may have an impact on the fibroblast component of the tumour microenvironment, potentially influencing SDF-1 and HGF expression which in turn could influence tumour progression. [ABSTRACT FROM AUTHOR]

This document is a correction notice for an article titled "YangZheng XiaoJi exerts anti-tumour growth effects by antagonising the effects of HGF and its receptor, cMET, in human lung cancer cells." The correction states that the conflict of interest declaration in the original article was incomplete, as several co-authors are employees of Yiling Medical Research Institute, a subsidiary of Yiling Pharmaceuticals. The publisher, Springer Nature, remains neutral regarding jurisdictional claims and institutional affiliations. The authors of the article are listed as Wen G. Jiang, Lin Ye, Fiona Ruge, Sioned Owen, Tracey Martin, Ping-Hui Sun, Andrew J. Sanders, Jane Lane, Lucy Satherley, Hoi P. Weeks, Yong Gao, Cong Wei, Yiling Wu, and Malcolm D. Mason. [Extracted from the article]

Tumour cell phagocytosis by antigen presenting cells (APCs) is critical to the generation of antitumour immunity. However, cancer cells can evade phagocytosis by upregulating anti-phagocytosis molecule CD47. Here, we show that CD47 blockade alone is inefficient in stimulating glioma cell phagocytosis. However, combining CD47 blockade with temozolomide results in a significant pro-phagocytosis effect due to the latter's ability to induce endoplasmic reticulum stress response. Increased tumour cell phagocytosis subsequently enhances antigen cross-presentation and activation of cyclic GMP-AMP synthase–stimulator of interferon genes (cGAS–STING) in APCs, resulting in more efficient T cell priming. This bridging of innate and adaptive responses inhibits glioma growth, but also activates immune checkpoint. Sequential administration of an anti-PD1 antibody overcomes this potential adaptive resistance. Together, these findings reveal a dynamic relationship between innate and adaptive immune regulation in tumours and support further investigation of phagocytosis modulation as a strategy to enhance cancer immunotherapy responses. Professional antigen presenting cells (APCs) are deterred from phagocytosing cancer cells that express CD47. Here, the authors show that in glioblastoma mouse models, temozolomide improves the phagocytosis effect of CD47 blockade in APCs and results in the activation of adaptive anti-tumour responses. [ABSTRACT FROM AUTHOR]

Background/Aim: Gremlin1 (GREM1) plays an important role in certain malignancies by antagonising bone morphogenetic proteins and regulating angiogenesis directly/indirectly. The present study aimed to investigate the role of Gremlin1 in the development and progression of gastric cancer (GC). Materials and Methods: Expression of GREM1 in GCs was examined using quantitative real time PCR and The Cancer Genomic Atlas (TCGA) data. Influence on cellular functions was determined in both Gremlin1 knockdown and overexpression cell line models. Results: GREM1 expression was up-regulated in GCs, which was correlated with poorer survival. Increased GREM1 expression was significantly correlated with tumour growth/invasion and lymphatic metastasis. Gremlin1 promoted proliferation and tumourigenic capacity of GC cells in vitro. GREM1 expression was associated with epithelial mesenchymal transition (EMT), angiogenesis and lymphangiogenesis in GC. Conclusion: Increased GREM1 expression in GCs is associated with disease progression and poor prognosis in which EMT, angiogenesis and lymphangiogenesis are likely involved. [ABSTRACT FROM AUTHOR]

Purpose: Receptor-like protein tyrosine phosphatase kappa (PTPRK) has been shown to exhibit homophilic binding. It is a putative tumour suppressor in primary central nervous system lymphomas and colorectal cancer. The present study investigated the expression of PTPRK in breast cancer and the biological impact of PTPRK on breast cancer cells. Methods: Expression of PTPRK protein and gene transcript was examined in a cohort of breast cancer patients. The association of PTPRK transcript level and pathological and clinical aspects was then analysed. Knockdown of PTPRK in breast cancer cells was performed using a specific anti-PTPRK transgene. The impact of PTPRK knockdown on breast cancer cells was investigated using in vitro cell function assays. Results: Lower levels of PTPRK transcripts were seen in the advanced breast cancer. The reduced PTPRK transcript levels were associated with poor prognosis of the disease. PTPRK transcript levels were decreased in the primary tumours of patients who died from breast cancer or had metastases. Patients with lower expression of PTPRK had shorter survival compared with those higher expression levels of PTPRK. Knockdown of PTPRK resulted in increased proliferation, adhesion, invasion, and migration of breast cancer cells in vitro. Conclusions: Decreased expression of PTPRK in breast cancer is correlated with poor prognosis. PTPRK is a negative regulator of adhesion, invasion, migration, and proliferation of breast cancer cells. This suggests that PTPRK is a potential tumour suppressor in breast cancer. [ABSTRACT FROM AUTHOR]

Wnt5a is a non-canonical signaling Wnt. Low expression of WNT5A is correlated with poor prognosis in breast cancer patients. The highly invasive breast cancer cell lines, MDA-MB-231 and 4T1, express very low levels of WNT5A. To determine if enhanced expression of WNT5A would affect metastatic behavior, we generated WNT5A expressing cells from the 4T1 and MDA-MB-231 parental cell lines. WNT5A expressing cells demonstrated cobblestone morphology and reduced in vitro migration relative to controls. Cell growth was not altered. Metastasis to the lung via tail vein injection was reduced in the 4T1-WNT5A expressing cells relative to 4T1-vector controls. To determine the mechanism of WNT5A action on metastasis, we performed microarray and whole-transcriptome sequence analysis (RNA-seq) to compare gene expression in 4T1-WNT5A and 4T1-vector cells. Analysis indicated highly significant alterations in expression of genes associated with cellular movement. Down-regulation of a subset of these genes, Mmp13, Nos2, Il1a, Cxcl2, and Lamb3, in WNT5A expressing cells was verified by semi-quantitative RT-PCR. Significant differences in transcript splicing were also detected in cell movement associated genes including Cd44. Cd44 is an adhesion molecule with a complex genome structure. Variable exon usage is associated with metastatic phenotype. Alternative spicing of Cd44 in WNT5A expressing cells was confirmed using RT-PCR. We conclude that WNT5A inhibits metastasis through down-regulation of multiple cell movement pathways by regulating transcript levels and splicing of key genes like Cd44. [ABSTRACT FROM AUTHOR]

Background: Psoriasin (S100A7) is a member of the S100 gene family. Alteration of Psoriasin expression has previously been reported to play an important role in cancer aggressive behaviour. The current study sought to investigate the level of Psoriasin expression at the mRNA level in a cohort of patients with non-small cell lung cancer (NSCLC), the association with clinical implication and outcomes, and the molecular and cellular impact of the protein on lung cancer cells. Methods: Fresh frozen NSCLC cell carcinoma tissues, along with matched normal tissues were obtained from 83 NSCLC patients who received curative resection from January 2003 to December 2011. The expression of Psoriasin in the NSCLC specimens was assessed using both quantitative real time PCR (QPCR) and immunochemical staining. Knockdown and forced expression of Psoriasin in NSCLC cell lines were carried out using constructed plasmid vectors carrying either ribozyme transgenes targeting human Psoriasin or full-length coding sequence, respectively. The effect of Psoriasin on the functions of NSCLC cells was determined using a variety of in vitro cell function assays. Results: Higher mRNA levels of Psoriasin were observed in tumour tissues when compared to both the paired normal background tissues and none paired normal tissues (p = 0.0251 and 0.0195). The mRNA level of Psoriasin was found to be higher in the squamous carcinoma (P=0.035). Higher Psoriasin expression is associated with poor prognosis. The cell function tests had supportive results to the clinical findings. Over-expression of Posriasin in lung cancer cells (SK-MES-1) resulted in an increase in in vitro growth and invasiveness. In contrast, Psoriasin knockdown suppressed cell growth and invasion (P<0.05), but increased cell adhesion (P<0.05). Conclusions: Psoriasin expression is increased in lung cancer, more specifically in lung squamous carcinoma compared with adenocarcinoma, and is associated with poor prognosis. Psoriasin plays crucial roles in regulating the growth and invasion of lung cancer cells. [ABSTRACT FROM AUTHOR]

Background: Metastasis suppressor-1 (MTSS1) has been proposed to function as a cytoskeletal protein with a role in cancer metastasis. Recent studies have demonstrated the clinical significance of MTSS1 in certain type of cancers, yet the clinical relevance of MTSS1 in oesophageal squamous cell carcinoma (ESCC) has not been reported.Methods: In this study, we assessed the expression levels of MTSS1 in tumours and its matched adjacent non-tumour tissues obtained from 105 ESCC patients. We also used ESCC cells with differing MTSS1 expression and assessed the influence of MTSS1 on ESCC cells.Results: Down-regulation of MTSS1 expression was observed both in oesophageal tumour tissues and ESCC cancer cell lines. We also reported that MTSS1 expression was associated with tumour grade (p = 0.024), lymph node metastasis (p = 0.010) and overall survival (p = 0.035). Patients with high levels of MTSS1 transcripts had a favorable prognosis in comparison with those who had reduced or absent expression levels. Using over-expression and knockdown approach, we created sublines from ESCC cells and further demonstrated that MTSS1 expression in ESCC cells significantly influenced the aggressiveness of the oesophageal cancer cells, by reducing their cellular migration and in vitro invasiveness.Conclusion: MTSS1 serves as a potential prognostic indicator in human ESCC and may be an important target for cancer therapy. [ABSTRACT FROM AUTHOR]

Background: EPLIN (epithelial protein lost in neoplasm) is a cytoskeletal associated protein involved in the regulation of actin dynamics and subsequently in cell motility. EPLIN expression is frequently reduced in a variety of cancer cells and tissues and this loss may account for increased invasive traits in cancer cells. The current study aimed to assess the role of EPLIN in endothelial cells and the angiogenic process. Methods: EPLINα expression was enhanced in HECV endothelial cells through transfection with a pEF6 expression plasmid containing the full coding sequence of EPLINα. The impact of EPLINα on HECV cells was then assessed using a range of in vitro and in vivo models. Results: Transfection and over-expression of HECV cells with EPLINα expression plasmid resulted in a significant reduction in cell-matrix adhesion ( P = 0.003), the rate of migration ( P = 0.009) and tubule formation ( P = 0.007) and also enhanced paxillin staining compared to transfection controls. Additionally, MDA-MB-231 breast cancer cells co-injected with HECV cells developed at a slower rate than those co-injected with HECV cells (day 26, P = 0.05; day 33, P = 0.065, overall deviation over time P < 0.001). Treatment of HECV cells with ERK inhibitor could rescue HECV cells from the inhibitory effect of EPLINα over-expression on tubule formation, returning these cells to control levels. Conclusions: EPLINα over-expression can regulate HECV cell motility, matrix adhesion and tubule formation in vitro and slow in vivo tumour formation, suggesting an anti-angiogenic role for EPLINα. Extracellular signal-regulated kinase (ERK) also appears to be linked to the ability of EPLINα to inhibit tubule formation in vitro. [ABSTRACT FROM AUTHOR]

Background: iASPP is a key inhibitor of tumour suppressor p53 and is found to be up-regulated in certain malignant conditions. The present study investigated the expression of iASPP in clinical lung cancer, a leading cancer type in the world, and the biological impact of this molecule on lung cancer cells. Methods: iASPP protein levels in lung cancer tissues were evaluated using an immunohistochemical method. In vitro, iASPP gene expression was suppressed with a lentvirus-mediated shRNA method and the biological impact after knocking down iASSP on lung cancer cell lines was investigated in connection with the p53 expression status. Results: We showed here that the expression of iASPP was significantly higher in lung cancer tissues compared with the adjacent normal tissues. iASPP shRNA treatment resulted in a down-regulation of iASPP in lung cancer cells. There was a subsequent reduction of cell proliferation of the two lung tumour cell lines A459 and 95D both of which had wild-type p53 expression. In contrast, reduction of iASPP in H1229 cells, a cell with little p53 expression, had no impact on its growth rate. Conclusions: iASPP regulates the proliferation and motility of lung cancer cells. This effect is intimately associated with the p53 pathway. Together with the pattern of the over-expression in clinical lung cancers, it is concluded that iASPP plays an pivotal role in the progression of lung cancer and is a potential target for lung cancer therapy. [ABSTRACT FROM AUTHOR]

Background and aims: Bone morphogenetic proteins (BMPs) have a diverse role and they act in a time, concentration and cell type specific manner. They regulate cellular motility and the cells ability to invade. Recently, BMP molecules have further been shown to have an impact on the biological behaviour of breast cancer cells. In this study, we looked, for the first time, at the expression of a panel of BMPs in breast carcinomas. Method: Fresh frozen primary human breast cancer tissues (n-120) and non-neoplastic mammary tissues (n= 32) were used. The distribution and location of BMPs was assessed using immunohistochemical methods and the transcript levels of BMPs (BMP-1, -2, -3, -4, -5, -6, and -7) were determined using quantitative RT-PCR. The results were analysed against the clinical, pathological and follow-up (10 years) data. Results: BMP-2 and BMP-7 exhibited contrasting patterns of expression in normal and tumour tissues, wherein BMP-2 transcript level was lower and BMP-7 was higher in breast tumours than normal tissues. BMP-2 transcript was also significantly lower in tumours from patients with a moderate and poor prognosis than from those with a good prognosis (p=0.04). BMP-2 and BMP-7 also showed a significant difference between node positive and node negative tumours (p =0.033 and p=0.031 respectively). BMPs 1, 3, 4, 5 and 6 showed an inconsistent variation in transcript levels within the cancer subgroups with no statistically significant results. Conclusion: This study has demonstrated a differential pattern of expression of BMP molecules in breast cancer and reveals a potential prognostic value of BMP-2 and BMP-7 for patients. The findings also suggest that these BMPs may be potential therapeutic targets. [ABSTRACT FROM AUTHOR]

Purpose: WAVE1 belongs to the Wiskott-Aldrich syndrome family of proteins, which have an integral part in cell motility, a crucial step in cancer metastasis. We investigated the expression pattern and the effects of manipulating endogenous WAVE1 expression in prostate cancer cells. Materials and Methods: WAVE1 protein expression in normal and cancer specimens, and in prostate cell lines was assessed using immunohistochemistry and reverse transcriptase-polymerase chain reaction, respectively. Hammerhead ribozyme transgenes were synthesized and cloned into the mammalian expression vector pEF6/V5-His TOPO TA, and transfected by electroporation into PC-3ΔW1R1/2 and DU-145ΔW1R1/2 cell lines. In vitro invasion, adhesion and growth assays were used to assess the impact of WAVE1 knockdown. Results: Immunohistochemistry of prostate tissue specimens showed that the cytoplasm of cancer cells had stronger staining than normal epithelium. Reverse transcriptase-polymerase chain reaction for WAVE1 showed strong expression in the PC-3 (European Collection of Cell Cultures, Salisbury, United Kingdom) and DU-145 (ATCC®) cell lines. WAVE1 knockdown was associated with a significant decrease in invasion but not in adhesion. The mean ± SEM number of invading PC-3ΔW1R1 and PC-3ΔW1R2 cells was 7.27 ± 0.38 and 6 ± 0.29, respectively, compared to 12.27 ± 0.42 PC-3WT cells (p <0.001). Similarly the mean number of invading DU-145ΔW1R1 and DU-145ΔW1R2 cells was 9.20 ± 0.70 and 11.60 ± 0.84 compared to 14.80 ± 0.24 DU-145WT cells (p <0.001). Conclusions: To our knowledge this is the first report of the expression pattern of WAVE1 in prostate cancer cell lines and tissues, and the functional impact of WAVE1 knockdown. Further investigations to assess WAVE1 as a potential target for anti-metastasis therapy must be explored. [Copyright &y& Elsevier]

Phosphatidylinositol transfer protein membrane-associated 1 (PITPNM1) contains a highly conserved phosphatidylinositol transfer domain which is involved in phosphoinositide trafficking and signaling transduction under physiological conditions. However, the functional role of PITPNM1 in cancer progression remains unknown. Here, by integrating datasets of The Cancer Genome Atlas (TCGA) and Molecular Taxonomy of Breast Cancer (METABRIC), we found that the expression of PITPNM1 is much higher in breast cancer tissues than in normal breast tissues, and a high expression of PITPNM1 predicts a poor prognosis for breast cancer patients. Through gene set variation analysis (GSEA) and gene ontology (GO) analysis, we found PITPNM1 is mainly associated with carcinogenesis and cell-to-cell signaling ontology. Silencing of PITPNM1, in vitro, significantly abrogates proliferation and colony formation of breast cancer cells. Collectively, PITPNM1 is an important prognostic indicator and a potential therapeutic target for breast cancer. [ABSTRACT FROM AUTHOR]

The cellular function and the role of matriptase-2 in cancer progression are poorly understood. This study assesses the importance of this protease in prostate cancer cell lines. Two prostate cancer cell lines, PC-3 and DU-145, previously displaying minimal expression of matriptase-2, were forced to over-express matriptase-2 using a human mammalian expression construct. Over-expression of matriptase-2 significantly reduced the invasive capacity and significantly slowed the migration rates of PC-3 and DU-145 cells in vitro. Similarly, PC-3 cells containing the matriptase-2 expression plasmid were dramatically less able to survive, grow and develop into noticeable tumours, compared to control PC-3 cells containing an empty plasmid alone, following subcutaneous inoculation into CD1 nude mice. This trend was observed throughout the experiment, becoming apparent after the initial reading on day 7 (P = 0.0002) and continuing to the experimental end point at day 27 (P = 0.0002). Enhanced matriptase-2 levels were also seen to correlate with increased fluorescent staining of the paxillin and FAK adhesion molecules, where a greater extent of these molecules were localised to the focal adhesion complexes. This data suggests a suppressive role for matriptase-2 in the invasion and migration of prostate cancer cells in vitro and also in their development and growth in vivo, highlighting the potential of this molecule to interfere with key stages of metastasis. Furthermore, the data presented implies a possible connection between matriptase-2 and the paxillin and FAK adhesion molecules which may ultimately contribute to the reduced migration rates seen in this study. J. Cell. Physiol. 216: 780–789, 2008, © 2008 Wiley-Liss, Inc. [ABSTRACT FROM AUTHOR]

Introduction: To investigate the expression of EPLIN-α, epithelial protein lost in neoplasm, in human breast cancer tissues/cells and investigate the cellular impact of EPLIN-α on breast cancer cells. Experimental design: EPLIN-α was determined in tumour (n = 120) and normal mammary tissues (n = 32), and cancer cell lines (n = 16). Cell invasion, in vitro and in vivo growth of cells transfected with EPLIN-α were evaluated using in vitro invasion assay, in vitro and in vivo tumour model. Cellular migration was analysed using Electric Cell Impedance Sensing assays. Results: Low level of EPLIN-α was seen in tumour tissues. Grade-2/3 tumours had significantly lower levels of EPLIN-α compared with grade-1 (p = 0.047 and p = 0.046 vs grade-1, respectively). Patients with poor prognosis had a significantly lower levels of EPLIN-α compared with those with good prognosis (p = 0.0081). Patients who developed recurrence and died of breast cancer had significantly lower levels of EPLIN-α compared with those who remained disease free (p = 0.0003 and p = 0.0008, respectively) (median follow-up 10 years). Patients with high levels of EPLIN-α transcript had a longer survival than those with low levels. Over-expression of EPLIN-α in breast cancer cells by way of transfection rendered cells less invasive, less motile and growing at a slower pace in vitro and in vivo. An ERK inhibitor was shown to be able to abolish the effect of EPLIN expression. Conclusion: It is concluded that expression of EPLIN-α in breast cancer is down-regulated in breast cancer cells and tissues, a change linked to the prognosis. EPLIN-α acts as a potential tumour suppressor by inhibition of growth and migration of cancer cells. [ABSTRACT FROM AUTHOR]

Purpose: We investigated the effect of manipulating endogenous BMP-7 expression on the invasion and motility of prostate cancer cells and the resulting effect on its antagonists using a ribozyme transgene. Materials and Methods: A hammerhead ribozyme transgene was synthesized and cloned into a mammalian expression vector (pcDNA3.1/nt-GFP-TOPO). PC-3 cells (American Type Culture Collection, Manassas, Virginia) were transfected with the ribozyme transgene (PC-3 ΔBMP7) or with an empty plasmid (PC-3pcDNA/GFP) by electroporation. Invasion and motility were accessed by in vitro invasion and motility assays. Results: The ribozyme decreased BMP-7 expression at the mRNA and protein levels in PC-3 cells. Invasive potential was significantly increased following the loss of BMP-7 expression. The mean ± SD invading cell number for PC-3 ΔBMP7 was 231.3 ± 28.6 vs 7.1 ± 4.4 for the WT cell line PC-3WT and 2.7 ± 2 for PC-3pcDNA/GFP (each p <0.001). BMP-7 knockdown in PC-3 cells significantly increased motility with a migrating cell number for PC-3 ΔBMP7 of 24 ± 7.5 compared with 10.2 ± 4.5 for PC-3WT and 11.3 ± 7.5 for PC-3pcDNA/GFP (p <0.01 and 0.011, respectively). The change in motility was seen together with changes in the cellular location of paxillin and focal adhesion kinase (p125FAK). Interestingly the loss of BMP-7 resulted in decreased noggin and follistatin expression. Conclusions: The loss of endogenous BMP-7 from prostate cancer cells is associated with increased invasiveness and motility, which appears to be facilitated by changes in the level of the BMP antagonists noggin and follistatin. Endogenous BMP-7 has an important role in controlling noggin and follistatin expression. [Copyright &y& Elsevier]

Background. Prostate transglutaminase (TGase-4), has been identified as a TGase that has a pattern of prostate specific expression. This study sought to determine (i) the level of TGase-4 in human cancer cell lines including prostate cancer cells; (ii) to investigate the effect after knocking down the expression of TGase-4 transcript using a ribozyme transgene; (iii) to establish the effect of HGF/SF, a pro-invasion/metastasis cytokine, on the invasive capacity of TGase-4 knockdown ceils. Methods: RT-PCR and quantitative RT-PCR were used to assess the presence of TGase-4 transcript at the mRNA level in a panel of 26 cell lines including 6 prostate cancer cell lines (PC-3, DU-145, CA-HPV-10, PZ-HPV-7, PNT2C2 and PNT1A). A ribozyme transgene consisting of hammerhead ribozyme and antisense specific to TGase-4 was constructed using a pEF6 expression vector and transfected into CA-HPV-10 cell, a cell line highly expressed TGase-4. An in vitro invasion assay assessed the effects of HGF/SF on cell invasiveness. Results: TGase-4 transcript was detected in a number of cell lines, including prostate, colorectal, lung and breast tumour cells. At the mRNA level, TGase-4 was strongly expressed in CA-HPV-10 cells with lower levels of expression seen in PC-3, DU-145, PZ-HPV-7 and PNT1A cells. TGase-4 knock-down in CA-HPV-10 cells was established at the mRNA level. HGF/SF significantly increased the invasion of wild type (21.67 ± 0.88; P<0.001 vs. control 8.67 ± 0.67) and control plasmid transfected cells (16.33 ± 0.88; P<0.001 vs. control 7.67 ± 0.88) compared with untreated cells. However, cells transfected with the TGase-4 ribozyme transgene had a reduced invasive capacity (9.33 ± 0.88 P<0.01 vs. control 4.33 ± 0.88) through Matrigel. Conclusion: TGase-4 has a relatively wide profile of expression in human cancer cell lines and is strongly expressed in the low invasive CA-HPV-10 prostate cancer cell line. The molecule is associated with the invasive potential of prostate cancer cells. [ABSTRACT FROM AUTHOR]

Purpose: Matriptase-1 has been implicated as playing an important role in various types of cancer progression through many different cancer related pathways. In the current study we assessed the efficacy of targeting matriptase-1 using ribozyme technology in vitro and in vivo. Experimental Design: Matriptase-1 expression was reduced in the PC-3 and DU-145 cell line using hammerhead ribozyme transgenes. In vitro assays were set up to assess changes in growth, invasion, adhesion and migration in these cells. In vivo tumour development model was also used to examine the efficacy of targeting matriptase-1 in a living environment. Results: The in vitro results suggest an overall reduction in the aggressive nature of the two cell lines (PC-3 and DU-145) when matriptase-1 levels are reduced, with properties such as growth, invasiveness and migration all being reduced (in most cases a greater than 50% reduction in migration and invasion compared to the control was observed), though strangely an increase in adhesion is seen in the PC-3 knockout. The in vitro data is strongly backed up by the results of the in vivo work which demonstrates matriptase-1 deficient cells have a substantially reduced ability to grow and develop in vivo compared to control cells when explanted into nude mice, with significant differences in growth and development (P ≤ 0.05) being seen after 7 days, and highly significant differences (p ≤0.001) after 15 days. Conclusions: Together this data strongly implicates matriptase-1 as playing a vital role in the aggressive nature and progression of prostate cancer. [ABSTRACT FROM AUTHOR]

Abstract: 5-lipoxygenase (5-LOX)-activating protein, 5-LOXAP also known as LOX5AP or FLAP, is a protein that works closely with 5-LOX in regulating the metabolism of arachidonate. Some of the eicosanoid products of 5-LOX/5-LOXAP are known to play active roles in the function of cancer cells, including breast cancer cells. The current study investigated the expression of 5-LOXAP in clinical breast cancer and the prognostic impact of 5-LOXAP and 5-LOX in patients with breast cancer. A cohort of breast tumour tissues () with normal background tissues () were investigated. 5-LOXAP and 5-LOX transcripts were determined using RT-PCR and quantitative RT-PCR. Levels of the transcripts were analysed against clinical and pathological information. Breast tumour tissues had significantly higher levels of 5-LOX transcript compared with normal tissues (). The transcript was seen at significantly higher levels in node positive tumours than that in node negative tumours (). The prognostic significance was assessed using both a prognostic index and clinical outcome. Value of 5-LOXAP was first demonstrated when using the Nottingham Prognostic Index (NPI) as an indicator, in that patients with predicted poor prognosis had significantly higher levels of 5-LOXAP than patients with good prognosis (). Furthermore, patients who died of breast cancer-related causes had significant higher levels of 5-LOXAP than those patients who remained disease free, following a median 10-year followup. A survival analysis has shown that high levels of 5-LOXAP were significantly correlated with overall survival (mean survival 109.6 month vs. 139.4 months, in tumour from patients with high and low levels of 5-LOXAP, ). The same disadvantage of high levels of 5-LOXAP was also seen with disease-free survival (105.2 months vs. 135.6 months, ). Analysis of 5-LOXAP together with 5-LOX transcript did not enhance the significance of the survival. However, when 5-LOXAP was considered together with 12-LOX, it improved the predictive power for both overall and disease-free survival (109.0 month vs. 143.1 months, for overall survival and 98.3 months vs. 141.3 months for disease-free survival, ). In conclusion, 5-LOXAP expression was aberrant in human breast cancer, particularly in aggressive tumours. Furthermore, 5-LOXAP had a significant prognostic value in patients with breast cancer. This identifies 5-LOXAP as a potential therapeutic target in breast cancer. [Copyright &y& Elsevier]

Abstract: Background: Com-1 is a molecule that has recently discovered to have putative action on the metastatic nature of cancer cells. The current study investigated the impact of Com-1 on oestrogen regulated cell growth of breast cancer cells and explored the potential link between Com-1 and ER-β. Method: Full length Com-1 cDNA was isolated from normal mammary tissues. Ribozyme transgenes that specifically targeted human Com-1 were constructed using the pEF6/V5-His vector. Expression of Com-1 was assessed at both mRNA and protein levels. Interaction of Com-1 with other candidate molecules was studied using immunoprecipitation and Western blotting. Results: Elimination of Com-1 by way of ribozyme transgenes results in breast cancer cells with increased rate of growth and increased invasive potential. In contrast, over-expression of Com-1 in the cancer cells had an opposite effect. In ER-α-negative/ER-β positive MDA MB-231 cells, elimination of Com-1 caused more vigorous growth in response to 17-β-estradiol. However, the effect of Com-1 modification on MCF-7, which is positive for both ER-α and ER-β, was less clear. Protein interaction analysis has indicated that the Com-1 and ER-β were mutually co-precipitated with each other in breast cancer cells. Immunocytochemical staining revealed that Com-1 was primarily present in the nucleus, with some degree of cytoplasmic staining, and that the distribution of Com-1 was identical to that of ER-β. 17-β-Estradiol stimulation resulted in reduction of nucleic staining of Com-1. This reduction of nucleic Com-1 can be reverted when ubiquitin inhibitor, ubiquitin aldehyde or the lactacystin proteosome inhibitor was present, suggesting a pivotal role of the ubiquitin–proteosome pathway in the Com-1/ER-β complex. Conclusion: Com-1 plays a tumour suppressor role in breast cancer cells and is involved in oestrogen-regulated cell growth. This action is potentially exerted by interacting with ER-β, in human breast cancer cells. The fate of Com-1 can be dually regulated by oestrogen and ubiquitin pathway. [Copyright &y& Elsevier]

Abstract: SPARC (secreted protein acidic and rich in cystein), also known as osteonectin and BM40, is a secreted glycoprotein. It confers matrix adhesion of cells including cancer cells, thus promoting cell migration. SPARC has been shown to be involved in the aggressive nature of cancer. The current study investigated the role of a n-6 polyunsaturated fatty acid, gamma linolenic acid (GLA) on the expression and secretion of SPARC from cancer cells. Human breast cancer cell line MCF-7 and MDA-MB-231, human colon cancer cells HT115 and HRT-18 were used in the study. Cancer cells were treated with GLA or other fatty acids over a range of concentrations. Presence of SPARC in the supernatant and in the cell lysate were analysed using Western blotting. Cellular SPARC was also assessed using immunocytochemistry. SPARC transcript in these cells were studied using RT-PCR. Cell–matrix adhesion was determined using a cell–matrix adhesion assay and cell migration analysis. Treatment of MDA-MB-231 and HT115 cells with GLA, at non-toxic levels, resulted in reduction of SPARC in supernatant as well as in the cell lysate. In contrast, there were little changes in the supernatant SPARC in MCF-7 and in HRT-18 cells. Cellular SPARC, as revealed by immunocytochemistry, also demonstrated a similar trend of changes as seen with protein blotting. Analysis of the SPARC transcript using RT-PCR has shown an up-regulation of SPARC mRNA by the fatty acid. GLA reduced cell–matrix adhesion in these cancer cells. It is concluded that GLA is a regulator of SPARC secretion and expression in cancer cells. It reduces the secretion of SPARC into surrounding environment, which may contribute to the reduction of cancer cells adhesion to the extracellular matrix and cell motility. [Copyright &y& Elsevier]

Simple Summary: Prostate cancer frequently metastasizes to the bone, where it forms primarily osteoblastic lesions. Currently there is no real therapeutic option for this late stage of disease, and understanding prostate cancer-bone interaction and communication is vital. Using a simple in vitro model of os-teoblast differentiation and mineralization, we studied this interaction and observed that prostate cancer cells secreted large quantities of extracellular vesicles containing microRNAs. When ex-posed to the extracellular vesicles, increased osteoblast differentiation and mineralization could be observed, and upon RNA-seq several of these microRNAs were implicated as upstream regulators of the mineralization process. These microRNAs also correlated with poor survival in online analysis of patient datasets. We characterized and validated four genes known to be targeted by microRNA-16, and found that extracellular vesicles could deliver miR-16, and increase minerali-zation. Skeletal metastases are the most common form of secondary tumour associated with prostate cancer (PCa). The aberrant function of bone cells neighbouring these tumours leads to the devel-opment of osteoblastic lesions. Communication between PCa cells and bone cells in bone envi-ronments governs both the formation/development of the associated lesion, and growth of the secondary tumour. Using osteoblasts as a model system, we observed that PCa cells and their conditioned medium could stimulate and increase mineralisation and osteoblasts' differentiation. Secreted factors within PCa-conditioned medium responsible for osteoblastic changes included small extracellular vesicles (sEVs), which were sufficient to drive osteoblastogenesis. Using MiR-seq, we profiled the miRNA content of PCa sEVs, showing that miR-16-5p was highly ex-pressed. MiR-16 was subsequently higher in EV-treated 7F2 cells and a miR-16 mimic could also stimulate mineralisation. Next, using RNA-seq of extracellular vesicle (EV)-treated 7F2 cells, we observed a large degree of gene downregulation and an increased mineralisation. Ingenuity® Pathway Analysis (IPA®) revealed that miR-16-5p (and other miRs) was a likely upstream effec-tor. MiR-16-5p targets in 7F2 cells, possibly involved in osteoblastogenesis, were included for val-idation, namely AXIN2, PLSCR4, ADRB2 and DLL1. We then confirmed the targeting and dow-regulation of these genes by sEV miR-16-5p using luciferase UTR (untranslated region) reporters. Conversely, the overexpression of PLSCR4, ADRB2 and DLL1 lead to decreased osteoblastogene-sis. These results indicate that miR-16 is an inducer of osteoblastogenesis and is transmitted through prostate cancer-derived sEVs. The mechanism is a likely contributor towards the for-mation of osteoblastic lesions in metastatic PCa. [ABSTRACT FROM AUTHOR]

Osteopontin (OPN) is identified as a diagnostic and prognostic biomarker of tumor progression and metastasis. In non-small-cell lung cancer (NSCLC), the functions of OPN have not been well characterized. The current study sought to investigate the clinical implications of OPN expression in NSCLC and the role of OPN in the aggressiveness of the lung cancer cells. Using a proteomics approach, we identified that phospho-RON (p-RON) was one of the most remarkably up-regulated proteins in OPN-overexpressing cells. The levels of OPN and RON transcripts were unveiled as independent prognostic indicators of survival in NSCLC (p = 0.001). Higher levels of OPN, RON and p-RON proteins were observed in tumor tissues. Knock down of the OPN gene suppressed the migration and invasion abilities of the A549 lung cancer cells which endogenously expresses OPN. While ectopic expression of OPN in the SK-MES-1 lung cancer cells increased levels of cellular invasion and migration. In addition, these changes were accompanied by a phosphorylated activation of RON. Small-molecule inhibition of RON or siRNA silencing of RON significantly reduced OPN-induced migration and invasion of lung cancer cells and had an inhibitory effect on the OPN-mediated cell epithelial-mesenchymal transition. Our study suggests that in NSCLC, the aberrant expression of OPN can be considered as an independent survival indicator and is associated with disease progression. OPN plays a crucial role in promoting migration and invasion properties of lung cancer cells through its phosphorylation activation of the RON signaling pathway, implying its potential as a therapeutic target in the treatment of NSCLC. [ABSTRACT FROM AUTHOR]

A near-infrared fluorescent probe, CyAc, is synthesized for accurately diagnosing cancer in vivo by sequential detection of Cys and H+. CyAc can not only achieve a good distinction between normal cells and cancer cells, but also distinguish normal mice from tumor mice. [ABSTRACT FROM AUTHOR]

Onthe basis of the polarization-dependent absorption of graphene undertotal internal reflection, we designed a graphene-based optical refractiveindex sensor with high resolution of 1.7 × 10–8and sensitivity of 4.3 × 107mV/RIU, as well asan extensive dynamic range. This highly sensitive graphene opticalsensor enables label-free, live-cell, and highly accurate detectionof a small quantity of cancer cells among normal cells at the single-celllevel and the simultaneous detection and distinction of two cell lineswithout separation. It provides an accurate statistical distributionof normal and cancer cells with fewer cells. This facile and highlysensitive sensing refractive index may expand the practical applicationsof the biosensor. [ABSTRACT FROM AUTHOR]

Abstract: How mutations or dysfunction of CFTR may increase the risk of malignancies in various tissues remains an open question. Here we report the interaction between CFTR and an adherens junction molecule, AF-6/afadin, and its involvement in the development of colon cancer. We have found that CFTR and AF-6/afadin are co-localized at the cell–cell contacts and physically interact with each other in colon cancer cell lines. Knockdown of CFTR results in reduced epithelial tightness and enhanced malignancies, with increased degradation and reduced stability of AF-6/afadin protein. The enhanced invasive phenotype of CFTR-knockdown cells can be completely reversed by either AF-6/afadin over-expression or ERK inhibitor, indicating the involvement of AF-6/MAPK pathway. More interestingly, the expression levels of CFTR and AF-6/afadin are significantly downregulated in human colon cancer tissues and lower expression of CFTR and/or AF-6/afadin is correlated with poor prognosis of colon cancer patients. The present study has revealed a previously unrecognized interaction between CFTR and AF-6/afadin that is involved in the pathogenesis of colon cancer and indicated the potential of the two as novel markers of metastasis and prognostic predictors for human colon cancer. [Copyright &y& Elsevier]

Gemcitabineis a nucleoside analogue commonly used in cancer therapybut with limited efficacy due to a high susceptibility to cancer cellresistance. The addition of a phosphoramidate motif to the gemcitabinecan protect it against many of the key cancer resistance mechanisms.We have synthesized a series of gemcitabine phosphoramidate prodrugsand screened for cytostatic activity in a range of different tumorcell lines. Among the synthesized compounds, one in particular (NUC-1031, 6f) was shown to be potent in vitro. Importantly,compared with gemcitabine, 6factivation was significantlyless dependent on deoxycytidine kinase and on nucleoside transporters,and it was resistant to cytidine deaminase-mediated degradation. Moreover, 6fshowed a significant reduction in tumor volumes in vivoin pancreatic cancer xenografts. The ProTide 6fis now in clinical development with encouraging efficacysignals in a Phase I/II study, which strongly supports the ProTideapproach to generate promising new anticancer agents. [ABSTRACT FROM AUTHOR]

Dendritic cells (DCs) play a pivotal role in initiating antigen-specific tumor immunity. However, the abnormal function of DCs owing to the immunosuppressive tumor microenvironment (TME) and the insufficient number of tumor infiltrating DCs could promote immune tolerance and tumor immune escape. Thus, there is great potential to employ DCs to induce efficient antitumor immunity. In this paper, we developed intelligent DCs (iDCs), which consist of nanoparticles loaded with photothermal agents (IR-797) and coated with a mature DC membrane. The DC cell membrane on the surface of iDCs preserves the ability to present antigens and prime T cells. The iDCs can also enter the lymph node and stimulate T cells. The activated T cells reduced the expression of heat shock proteins (HSPs) in tumor cells, rendering them more sensitive to heat stress. Subsequently, we used mild photothermal therapy (42–45 °C) to induce immunogenic cell death and contribute to a synergistic antitumor effect. iDCs as a refined and precise system in combination with DC-based immunotherapy and thermal therapy can be stored long-term and on a large scale, so they can be applied in many patients. [ABSTRACT FROM AUTHOR]

Abstract: Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) has recently emerged as a cancer therapeutic agent because it is capable of preferentially inducing apoptosis in human cancer over normal cells. The majority of human pancreatic cancers, unfortunately, are resistant to TRAIL treatment. Here, we show that the inhibition of caspase-8 cleavage is the most upstream event in TRAIL resistance in pancreatic cancers. TRAIL treatment led to the cleavage of caspase-8 and downstream caspase-9, caspase-3, and DNA fragmentation factor 45 (DFF45) in TRAIL-sensitive pancreatic cancer cell lines (BXPC-3, PACA-2). This caspase-8-initiated caspase cascade, however, was inhibited in TRAIL-resistant pancreatic cancer cell lines (PANC-1, ASPC-1, CAPAN-1, CAPAN-2). The long and short forms of cellular Fas-associated death domain-like interleukin-1β-converting enzyme-inhibitory protein (c-FLIPL, c-FLIPS) were highly expressed in the TRAIL-resistant as compared to the sensitive cells; knockdown of c-FLIPL and c-FLIPS by a short hairpin RNA (shRNA) rendered the resistant cells sensitive to TRAIL-induced apoptosis through the cleavage of caspase-8 and activation of the mitochondrial pathway. Receptor-interacting protein (RIP) has been reported in TRAIL-induced activation of NF-κB and we show here that knockdown of RIP sensitized the resistant cells to TRAIL-induced apoptosis. These results indicate the role of c-FLIP and RIP in caspase-8 inhibition and thus TRAIL resistance. Treatment of the resistant cells with camptothecin, celecoxib and cisplatin resulted in the downregulation of c-FLIP and caused a synergistic apoptotic effect with TRAIL. These studies therefore suggest that combination treatment with chemotherapy can overcome TRAIL resistance and enhance TRAIL therapeutic efficacy in treating pancreatic cancers. [Copyright &y& Elsevier]

Abstract: Based on our wide ranging knowledge of phosphoramidate ProTides as anti-viral agents we have tuned the lead anti-cancer agent thymectacin in the ester and amino acid regions and revealed a substantial enhancement in in vitro potency versus colon and prostate cancer cell lines. Twelve analogues have been reported, with yields of 29–78%. The compounds are fully characterised and data clearly reveal the presence of two phosphate diastereoisomers, as expected, in roughly equi-molar proportions. The compounds were evaluated in tissue culture versus three different tumour cell lines, using thymectacin as the control. It is notable that minor structural modification of the parent phenyl methoxyalaninyl structure of thymectacin leads to significant enhancements in potency. In particular, replacement of the methyl ester moiety in the lead by a benzyl ester gave a 175-fold boost in potency versus colon cancer HT115. This derivative emerges as a low micromolar inhibitor of HT115 cells and a new lead for further optimisation. [Copyright &y& Elsevier]