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Background Despite lack of clinical therapy in acute kidney injury (AKI) or its progression to chronic kidney disease (CKD), administration of growth factors shows great potential in the treatment of renal repair and further fibrosis. At early phase of AKI, administration of exogenous fibroblast growth factor 2 (FGF2) protects against renal injury by inhibition of mitochondrial damage and inflammatory response. Here, we investigated whether this treatment attenuates the long-term renal interstitial fibrosis induced by I/R injury. Methods Unilateral renal ischemia-reperfusion (I/R) with contralateral nephrectomy was utilized as an in vivo model for AKI and subsequent CKD. Rats were randomly divided into four groups: Sham-operation group, I/R group, I/R-FGF2 group and FGF2-3D group. These groups were monitored for up to 2 months. Serum creatinine, inflammatory response and renal histopathology changes were detected to evaluate the role of FGF2 in AKI and followed renal interstitial fibrosis. Moreover, the expression of vimentin, α-SMA, CD31 and CD34 were examined. Results Two months after I/R injury, the severity of renal interstitial fibrosis was significantly attenuated in both of I/R-FGF2 group and FGF2-3D group, compared with I/R group. The protective effects of FGF2 administration were associated with the reduction of high-mobility group box 1 (HMGB1) mediated inflammatory response, the inhibition of TGF-β1/Smads signaling induced epithelial-mesenchymal transition and the maintenance of peritubular capillary structure. Conclusions A single dose of exogenous FGF2 administration 1 hour or 3 days after reperfusion inhibited renal fibrogenesis and thus blocked the transition of AKI to CKD. Our findings provided novel insight into the role of FGF signaling in AKI to CKD progression and underscored potential of FGF based therapy for this devastating disease.

Background: TMSG1/LASS2 is emerging as an effective tumor suppressor as a transmembrane protein since it has been first cloned. We for the first time found by accident in our previous study TMSG1/LASS2 could be located in the nucleus, and it contained Homeodomain and nuclear localization signal (NLS). We aimed to investigate whether TMSG1/LASS2 possessed the function of transcription factor and identify the target genes regulated by the transcriptional regulation function of TMSG1/LASS2.Methods: We constructed and transfected three variants containing different functional domains of TMSG1/LASS2 to human prostate cancer cell PC-3M-1E8. Immunofluorescence was performed to detect the cellular localization of the three variants and identified its functional domain mediating it entering the nucleus. Chromatin immunoprecipitation (ChIP) associated with high-throughput sequencing was performed to select the target genes binding to TMSG1/LASS2 protein. Then we investigated the transcriptional regulation effect of TMSG1/LASS2 on one of the target genes Neural Proliferation, Differentiation and Control 1 (NPDC1) by dual-luciferase reporter system, and examined the expression of NPDC1 upon the overexpression or knockdown of TMSG1/LASS2, as well as the cell proliferation ability, apoptosis and cell cycle. Results: Immunofluorescence result revealed the variants deleted Homeodomain or NLS could not enter the nucleus. Neural Proliferation, Differentiation and Control 1 (NPDC1) gene, whose promoter fragment was obtained from ChIP using TMSG1/LASS2 antibody for further study. Dual-Luciferase reporter system revealed TMSG1/LASS2, as well as the variant with the deletion of NLS could enhance the transcription of NPDC1 promoter rather than the variant deleted Homeodomain. RT-qPCR and Western blot results demonstrated the gene and protein levels of NPDC1 were significantly increased upon TMSG1/LASS2 overexpression other than the variants without Homeodomain or NLS. Moreover, the level of NPDC1 were markedly decreased upon TMSG/LASS2 knockdown. Furthermore, the full length TMSG1/LASS2 was more able to inhibit cell proliferation, invasive ability and promote the cell apoptosis than the variants without Homeodomain or NLS. Conclusions: TMSG1/LASS2 could enter the nucleus to play transcription factor role mediated by its Homeodomain and NLS. TMSG1/LASS2 could promote the transcription and subsequent expression of NPDC1, one of its target genes. TMSG-1/LASS2 might inhibit cell proliferation, invasive ability, promote cell apoptosis and G0/G1 cell cycle arrest partially mediated by its Homeodomain and NLS. We for the first time propose a new transcriptional regulation function of TMSG1/LASS2 in addition to the known tumor suppressor function as a transmembrane protein.

Oral squamous cell carcinoma (OSCC) is the most common malignancy of the oral cavity. In the tumor microenvironment, tumor-associated neutrophils (TANs) can promote tumor growth, invasion, and metastasis. The aim of our study was to explore the relationship between neutrophils infiltration and Chemerin expression in tumor cells, as well as their relationship with the clinicopathological parameters and clinical prognosis of 74 cases of OSCC. We also explored the role of the interaction between neutrophils and Chemerin in the functions of OSCC cells (Cal27, SCC9, and SCC15) in vitro. Our results showed that in OSCC, Chemerin over-expression may increase neutrophils infiltration in tumor tissues. Chemerin over-expression and neutrophils infiltration were the prognostic factors of poor clinical outcomes. Furthermore, we discovered that neutrophils promoted OSCC migration, invasion, and proliferation and EMT through Chemerin. Neutrophils activated JAK2/STAT3 signaling through Chemerin and then up-regulated its downstream signaling target genes, such as Phospho-Rb, E2F1, CyclinE1, and CyclinD1. Taken together, our results revealed that neutrophils and Chemerin are potentially involved in OSCC progression and metastasis. Neutrophils may promote the JAK2/STAT3 signaling pathway and EMT in OSCC cells through Chemerin.

Triple negative breast cancer (TNBC) displays higher heterogeneity, stronger invasiveness, higher risk of metastasis and poorer prognosis compared with major breast cancer subtypes. KIF3A, a member of the kinesin family of motor proteins, serves as a microtubule‐directed motor subunit and has been found to regulate early development, ciliogenesis and tumorigenesis. To explore the expression, regulation and mechanism of KIF3A in TNBC, 3 TNBC cell lines, 98 cases of primary TNBC and paired adjacent tissues were examined. Immunohistochemistry, real‐time PCR, western blot, flow cytometry, short hairpin RNA (shRNA) interference, 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide (MTT), colony formation techniques, transwell assays, scratch tests, and xenograft mice models were used. We found that KIF3A was overexpressed in TNBC and such high KIF3A expression was also associated with tumor recurrence and lymph node metastasis. Silencing of KIF3A suppressed TNBC cell proliferation by repressing the Rb‐E2F signaling pathway and inhibited migration and invasion by repressing epithelial‐mesenchymal transition. The tumor size was smaller and the number of lung metastatic nodules was lower in KIF3A depletion MDA‐MB‐231 cell xenograft mice than in the negative control group. In addition, KIF3A overexpression correlated with chemoresistance. These results suggested that high expression of KIF3A in TNBC was associated with the tumor progression and metastasis., KIF3A，a member of kinesin super family, was over‐expressed in triple negative breast cancer (TNBC) tissues and such high KIF3A expression was associated with tumor recurrence and lymph node. Silencing of KIF3A suppressed TNBC cells proliferation, migration and invasion by repressing the Rb‐E2F signaling and EMT. And the tumor size and lung metastatic nodules were less in KIF3A depletion xenograft mice.

Xuan Zhou, 1,* Huihui Ji, 2,* Dongxue Ye, 2 Hong Li, 1 Fen Liu, 2 Haiyan Li, 2 Jin Xu, 1 Yujun Li, 1 Fenggang Xiang 1, 2 1Department of Pathology, The Affiliated Hospital of Qingdao University, Qingdao 266000, People’s Republic of China; 2Department of Pathology, School of Basic Medicine, Qingdao University, Qingdao 266071, People’s Republic of China*These authors contributed equally to this workCorrespondence: Fenggang XiangDepartment of Pathology, The Affiliated Hospital of Qingdao University, 16 Jiangsu Road, Qingdao 266000, People’s Republic of ChinaEmail xiangfenggang0532@163.comIntroduction: The aim of the present study was to examine the expression of ATAD2 in gastric cancer (GC) specimens and to evaluate its correlation with clinicopathologic features, including survival of GC patients. The potential roles of ATAD2 in the GC cell proliferation, apoptosis, and tumour growth were further explored.Materials and Methods: Quantitative reverse transcription-polymerase chain reaction (qRT-PCR), Western blotting and immunohistochemistry (IHC) were applied to determine the mRNA and protein expression of ATAD2 in GC and corresponding adjacent non-tumourous specimens. The relationship between ATAD2 expression and clinicopathological features of GC patients was analysed. Kaplan-Meier analysis was performed to assess the prognostic value of ATAD2 expression levels. The proliferation, colony formation, apoptosis and tumorigenesis roles of ATAD2 were measured using in vitro and in vivo experiments.Results: The expression of ATAD2 mRNA and protein was overexpressed in GC tissues compared with corresponding adjacent non-tumourous tissues. ATAD2 expression was significantly correlated with tumour size, tumour differentiation, and clinical tumour-node-metastasis (TNM) stage. Patients with high ATAD2 expression were likely to experience significantly shorter postoperative overall survival (OS) and disease-free survival (DFS). Multivariate Cox analysis suggested ATAD2 as an independent variable for OS and DFS. Knockdown of ATAD2 significantly suppressed cell proliferation, colony formation in vitro and tumorigenicity in vivo. Cell cycle and apoptotic assays showed that the anti-proliferative effect of pLV-ATAD2 shRNA was mediated by arresting cells in the G1 phase and inducing cell apoptosis. Silencing of ATAD2 reduced the expression of cyclinD1, ppRb, E2F1 and cyclinE and upregulated the expression of cleaved-PARP and cleaved-Caspase 3.Conclusion: Our study indicated that ATAD2 plays an important role in the process of tumorigenesis and progression in GC, and it could serve as a novel prognostic biomarker and a therapeutic target for the treatment of GC patients.Keywords: ATAD2, gastric cancer, proliferation, apoptosis, prognosis

Breast cancer is the most common malignant tumors in women. Kinesin family member 3B (KIF3B) is a critical regulator in mitotic progression. The objective of this study was to explore the expression, regulation, and mechanism of KIF3B in 103 cases of breast cancer tissues, 35 metastatic lymph nodes and breast cancer cell lines, including MDA-MB-231, MDA-MB-453, T47D, and MCF-7. The results showed that KIF3B expression was up-regulated in breast cancer tissues and cell lines, and the expression level was correlated with tumor recurrence and lymph node metastasis, while knockdown of KIF3B suppressed cell proliferation, migration, and invasion both in vivo and in vitro. In addition, UALCAN analysis showed that KIF3B expression in breast cancer is increased, and the high expression of KIF3B in breast cancer is associated with poor prognosis. Furthermore, we found that silencing of KIF3B decreased the expression of Dvl2, phospho-GSK-3β, total and nucleus β-catenin, then subsequent down-regulation of Wnt/β-catenin signaling target genes such as CyclinD1, C-myc, MMP-2, MMP-7 and MMP-9 in breast cancer cells. In addition, KIF3B depletion inhibited epithelial mesenchymal transition (EMT) in breast cancer cells. Taken together, our results revealed that KIF3B is up-regulated in breast cancer which is potentially involved in breast cancer progression and metastasis. Silencing KIF3B might suppress the Wnt/β-catenin signaling pathway and EMT in breast cancer cells.

Carcinoembryonic antigen cell adhesion molecule 1 (CEACAM1), a transmembrane glycoprotein, has multiple functions. In tongue squamous cell carcinoma (TSCC), CEACAM1 overexpression is correlated with neutrophil infiltration, and both are associated with poor clinical outcomes. However, the mechanism underlying CEACAM1's effect on neutrophil function in TSCC remains unclear. We cocultured tongue carcinoma cells overexpressing CEACAM1‐4L, CEACAM1‐4S and differentiated HL‐60 cells. This significantly upregulated the expression of MMP‐9, interleukin 8, and VEGF‐A in the differentiated HL‐60 cells and downregulated the expression of TNF‐α, relative to vector and blank control groups (P

Cisplatin-based chemotherapy is the first-line treatment for non-small cell lung cancer (NSCLC), but drug resistance occurs in most patients, leading to treatment failure. Recent studies have shown that epithelial-mesenchymal transition (EMT) is associated with drug resistance. However, the underlying mechanism is not entirely clear. In this study, first we showed significant positive correlation between the expression of ERCCl and vimentin, and significant negative correlation between the ERCCl and E-cadherin in the neoadjuvant chemotherapy group and the simple surgery group. Second, we showed that cisplatin-resistant A549 cells (A549/DDP) acquire EMT phenotype with high expression of drug-resistant proteins, P-gp and ERCC1. Knockdown of TGF-β1 may reverse EMT and significantly reduce the expression of P-gp and ERCC1. Moreover, A549/DDP cells become more sensitive to cisplatin. In summary, our results globally confirm a molecular and phenotypic association between chemoresistance and EMT of resistant tumour cells under a histological and cellular level. More importantly, silence of TGF-β1 may enhance sensitivity to cisplatin of A549/DDP through inducing the reversal of EMT and inhibiting the expression of resistance-associated proteins. Hence, inhibition of TGF-β1 could be considered as an effective strategy for eliminating resistant lung cancer.

Breast cancer is the most common cause of death among women. KIF3C, a member of kinesin superfamily, functions as a motor protein involved in axonal transport in neuronal cells. To explore the expression, regulation and mechanism of KIF3C in breast cancer, 4 breast cancer cell lines and 93 cases of primary breast cancer and paired adjacent tissues were examined. Immunohistochemistry, Real Time Polymerase Chain Reaction (RT-PCR), Western blot, flow cytometry, short hairpin RNA (shRNA) interference, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), colony formation techniques and xenograft mice model were used. We found that KIF3C was over-expressed in breast cancer tissues and such high KIF3C expression was also associated with tumor recurrence and lymph node metastasis. Silencing of KIF3C by shRNA inhibited epithelial-mesenchymal transition and metastasis by inhibiting TGF-β signaling and suppressed breast cancer cell proliferation through inducing G2/M phase arrest. The tumor size was smaller and the number of lung metastatic nodules was less in KIF3C depletion MDA-MB-231 cell xenograft mice than in negative control group. These results suggested that high expression of KIF3C in breast cancer may be associated with the tumor progression and metastasis.

Primary intravascular large B cell lymphoma (IVLBCL) of the endometrium is extremely rare. So far, only 5 cases have been reported in the English literature. We now report a new case of endometrial IVLBCL which exhibited distinct clinicopathological characteristics and meaningful laboratory tests, and also review the literature. A 66-year-old woman showed symptoms of chronic cough and choking sensation for 4 months. Following three days of vaginal bleeding she presented for examination and diagnosis. The percentage of monocytes in the blood was double that of normal levels. There was a polyp in the endometrium, which showed a number of medium-large lymphoid cells in dilated capillaries. Immunohistochemically, the lymphoid cells were immunoreactive to CD20, CD79a, Mum-1 and Foxp-1 with 85% cells immunoreactive to Ki-67. IVLBCL of the endometrium is rare and the clinical diagnosis is very difficult. Unexplained fever of old people and abnormal laboratory tests such as obvious abnormal monocyte distribution in the blood should alert the clinical doctor to the possibility of IVLBCL. A correct diagnosis mainly depends on pathological tests and immunohistochemical labeling. The prognosis of IVLBCL is poor and few patients survive longer than one year.

Mitotic centromere-associated kinesin (MCAK) is a microtubule depolymerase indispensable for microtubule binding during spindle formation. The purpose of this study was to investigate the association of MCAK expression with squamous cell carcinoma of the oral tongue (SCCOT).Immunohistochemistry was used in 47 cases of SCCOT. Determination of proliferation and migratory capabilities was performed with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and Transwell chamber assay, respectively, on cells from the human tongue squamous cell carcinoma cell line Tca8113 that were transfected with MCAK small interfering RNA (siRNA).MCAK expression level in oral tongue cancer tissue is significantly higher (P.01) than that of corresponding normal tissue. In addition, high expression of MCAK is significantly associated with lymph node metastasis (P.05) and tumor staging (P.01). Moreover, gene silencing of MCAK suppresses proliferation and migration of Tca8113 cells (P.05; P.01).The expression of MCAK may be associated with the progression of SCCOT.

Gangliocytic paraganglioma (GP) is a rare histologic type of neuroendocrine tumors. We report a case of pulmonary GP in a 29-year-old male presenting with an asymptomatic endobronchial nodule. Grossly, the tumor showed a 4.0x3.8x3.5 cm well-defined nodule with yellowish cut surface. Microscopically, the tumor was composed of three distinct cellular types: epithelioid cells, ganglion-like cells and spindle cells. Meanwhile, transitional cells, having morphologic features between ganglion-like and epithelioid cells, were also presented. The epithelioid cells arranged in various morphologic architectures, including Zellballen, papillary, cystic and microcystic pattern. The epithelioid cells were positive for AE1/AE3, CAM 5.2, chromogranin A and synaptophysin. Ganglion-like cells showed immunoreactivity for chromogranin A and synaptophysin. A few ganglion-like cells were also positive for AE1/AE3 and/or CAM 5.2. The spindle cells were positive for S-100 protein and neurofilament. The transitional cells showed a similar immunohistochemical profile to the epithelioid cells. The authors believe stem cell theory is a reasonable explanation for the origin of GP. GP probably originate from some kind of mucosa associated stem cell which can differentiate into diverse cellular lineages.

It is well known that vascular endothelial growth factor (VEGF) and its receptors are closely related to tumor angiogenesis, but the exact relationship with patients' prognosis is unclear till now. The aim of this study is to explore the expression of VEGF and its receptors KDR, Flt1 in pulmonary carcinoma and their relationship with patients' prognosis.The expression of VEGF, KDR and Flt1 was examined immunohistochemically by PV-9000 method in 75 cases of pulmonary carcinoma with complete follow-up records.There was an extensive expression of VEGF, KDR and Flt1, mainly in the cytoplasm of tumor cells (TCs), fibroblasts (FBs), and endothelial cells. The distribution of VEGF, KDR and Flt1 was heterogeneous, mainly located at periphery of the tumor mass or necrosis. The positive rate of VEGF, KDR and Flt1 in the TCs was all significantly higher than that in the FBs (P0.01, P0.02, P0.02). Both in TCs and FBs, the positive expression of VEGF, KDR and Flt1 was related to the postoperative survival of patients (P0.01, P0.01, P0.01; P0.01, P0.01, P0.05). The survival time in patients with positive VEGF, KDR or Flt-1 in TCs was significantly lower than that in those with corresponding negative one respectively (P0.0001, P0.0005, P0.0005). There was a positive correlation between VEGF and Flt1 in TCs (P0.01), between VEGF in FBs and Flt1 in TCs (P0.01), and also between VEGF and KDR or Flt1 in FBs (P0.01, P0.01).VEGF may act as a considerable promoting growth factor on tumor cells via Flt1, mainly in autocrine and less in paracrine manner. VEGF, KDR and Flt1 may exert important roles in prognosis of patients with pulmonary carcinoma.

Neoadjuvant chemotherapy (NACT) plays an important role in systemic chemotherapy for non-small cell lung cancer (NSCLC). P-glycoprotein (P-gp), lung resistance related protein (LRP), multidrug resistance-associated protein (MRP) and glutathione S-transferase (GST-π) may be associated to drug resisitance to chemotherapy in NSCLC. The aim of this study is to investigate the expressions of P-gp, LRP, MRP and GST-π in samples from NSCLC patients before and after treatment with NACT, and their quantitative changes, so that to evaluate the influence of NACT on drug resistance to chemotherapy of NSCLC.Total 92 specimens from 72 cases of NSCLC, including 52 samples of surgery excision from non-NACT-treated patients and 20 paired samples before and after NACT from the same patient, were studied. The expression of P-gp, LRP, MRP and GST-π was detected with tissue chip technique and immunohistochemistry. The quantitative analysis was carried out by computer image analysis system..In the samples before NACT, the positive rate of P-gp, LRP, MRP, GST-π expression was 66.67% (48/72), 72.22% (52/72), 81.94% (59/72), 83.33% (60/72), respectively. The expressive intensity of P-gp, LRP and GST-π was significantly stronger in adenocarcinoma than that in squamous cell carcinoma (P0.05, P0.001, P0.001, respectively); there was no significant difference in the expression of MRP between adenocarcinoma and squamous cell carcinoma (P0.05). In samples after treatment with NACT, the expression of P-gp, GST-π demonstrated by average optical density (AOD) and integral optical density (IOD) were significantly higher (P0.05, P0.001 respectively) than that in biopsied samples taken before NACT; The change in expression of P-gp, GST-π was also showed difference by different histopathological types, differentiations, ages, sizes, clinical stages as well as lymph node metastasis or not (P0.05 or P0.001). There was no significant difference between samples taken before and after NACT (P0.05) in the expression of LRP and MRP demonstrated by both of AOD and IOD.The results suggest that drug resistance in adenocarcinoma is primarily stronger than that in squamous cell carcinoma. NACT may enhance acquired drug resistance of NSCLC through inducing the expression of drug resistance protein. The results indicate that acquired drug resistance must be considered with the application of NACT to NSCLC patient in clinic, especially to patient in stage I and II. Since NACT may lead to the enhancement of acquired drug resistance in stage I and II, this may dwindle the therapeutic effect of chemotherapy after surgery. Comparative examination of drug resistance proteins before and after NACT, combining with comprehensive consideration of chemical regimens of NACT, should be recommended during chemotherapy of NSCLC for both before and after surgery.

We studied the relationships among vascular endothelial growth factor (VEGF), its receptors (Flt1 and Flk1), and MIB-1. Their expression in 47 diffusely infiltrating astrocytomas obtained at surgery or autopsy was investigated by the ABC method and analyzed quantitatively. The positive rate of VEGF in tumor cells was higher than that in endothelial cells, and Flk1 was lower in tumor cells (P0.01, 0.01), whereas Flt1 in both tumor cells and endothelial cells was found at similar levels (P0.05). In tumor cells, VEGF became high with increased histological grades (P0.01). whereas both Flt1 and Flk1 were higher in grade 4 than in grades 2 and 3 (P0.01, 0.05). VEGF, Flt1, and Flk1 in endothelial cells were also highly expressed in grade 4 (P0.01). The distribution of MIB-1-positive nuclei in grade 4 was similar to VEGF, and the percent of positivity from grade 2 to grade 4 also increased (P0.01). There was a linear positive correlation between VEGF and both Flt1 and Flk1 in both tumor cells and endothelial cells (P0.01). So was the percent of positivity with VEGF, Flt1, and Flk1 in tumor cells and endothelial cells (P0.01). The experiment suggests that VEGF may act as a growth factor for both endothelial cells and tumor cells. VEGF, Flt1, and Flk1 can be considered as indicators of the malignancy potential of diffusely infiltrating astrocytomas. The expression of VEGF and the two receptors may be affected by the proliferative activity of tumor cells.