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Background: Platinum is widely used in the neoadjuvant and metastatic treatment of breast cancer, but increasingly drug resistance is the main cause of cancer recurrence and treatment failure. Our study aimed to investigate the mechanisms by which upregulation of estrogen-related receptor α (ERRα) induced chemoresistance in breast cancer. Methods: Immunohistochemistry (IHC) was used to determine the expression of ERRα in breast cancer and adjacent tissues. Functional analyses (in vitro and in vivo) were performed to confirm the role of ERRα in cancerogenesis and cisplatin chemoresistance in breast cancer. RNA-sequencing, ChIP and dual luciferaseassays were performed to identify the mechanisms by which ERRα promotes chemoresistance in breast cancer. Results: Analyses of ERRα expression among a case-control cohort of 63 annotated tumor specimens demonstrated that ERRα expression was highly expressed in breast cancer tissues. Overexpression of ERRα promoted cell proliferation and metastasis of breast cancer in vitro and in vivo, increased chemoresistant of cisplatin and enhanced the pluripotency; while ERRα knockdown resulted in the opposite effects. We show that ERRα can directly induce CCNE2 expression through binding its promoter region then enhanced the pluripotency of cancer cells and facilitated chemoresistance Overexpression of CCNE2 could reverse the sensitivity of breast cancer cells to cisplatin caused by ERRα depletion, thus resulting in accelerated tumor growth. Conclusions: Our study demonstrates that ERRα acts as oncogene in breast cancer and promotes cisplatin resistance by regulating the transcription of CCNE2 and may serve as a therapeutic target in breast cancer.

Vesicle amine transport-1 (VAT-1) was implicated in regulation of vesicular transport, mitochondrial fusion, axonal growth, phospholipid transport and cell migration. However, the role of VAT-1 in tumour biology and disease progression of hepatocellular carcinoma (HCC) remains unknown. Here, we first investigated the expression of VAT-1 in clinical HCC samples by immunohistochemistry and in various transcriptomic datasets by bioinformatics. The biological functions of VAT-1 in HCC were then explored by using a variety of techniques including in vitro cell-based assays, in vivo xenograft models, high throughput mRNA-Seq, KEGG pathway enrichment, flow-cytometry analysis, immunoassays and bioinformatics. Underlying mechanisms were further verified in clinical tumour specimens. We demonstrated that VAT-1 is significantly upregulated in tumour tissues and associated with tumour size, invasion, clinical stage and overall survival of patients. Multivariate Cox analysis indicated VAT-1 is an independent unfavourable prognostic factor. VAT-1 downregulation significantly inhibited cell growth and colony formation in vitro and xenograft growth by decreasing proliferation and increasing apoptosis. Mechanistic studies revealed VAT-1 downregulation inhibits tumour growth through induction of the cell cycle arrest at G1-G0 by regulating the expression of cyclin D1, cyclin D3, CDK6, c-Myc and MCL-1. Strikingly, VAT-1 regulates STAT3 phosphorylation at Y705, nuclear translocation of pSTAT3-Y705, EGF-induced STAT3 signalling and consequently the expression of downstream c-Myc and cyclin D1. In both established HCC cell lines and clinical tumour samples, VAT-1 regulates numerous pathways including cell cycle, ErbB pathway, EGFR tyrosine kinase inhibitor resistance and JAK-STAT pathway; VAT-1 expression is significantly correlated with core components of EGF-EGFR-STAT3-cell cycle axis, highlighting the role of VAT-1 in regulation of EGF-STAT3-c-Myc-cyclin D/CDK6 signalling. Our results provide new insights into the carcinogenesis and disease progression of HCC and rationales for the development of novel intervention strategies against HCC. VAT-1 could serve as an independent prognostic biomarker for predicting clinical outcome of HCC patients.

Background Chemotherapy is one of the most efficient therapeutic methods for cancer, but increasingly serious drug resistance is the main cause of cancer recurrence and treatment failure. Our study aimed to investigate the mechanisms by which upregulation of estrogen-related receptor α (ERRα) induced chemoresistance in breast cancer cells. Methods Immunohistochemistry (IHC) was used to determine the expression of ERRα in breast cancer and adjacent tissues. Functional analyses (in vitro and in vivo) were performed to confirm the role of ERRα in cancerogenesis and cisplatin chemoresistance in breast cancer. RNA-sequencing, ChIP-seq and dual luciferase assays were performed to identify the mechanisms by which ERRα promotes chemoresistance in breast cancer. Results ERRα was highly expressed in breast cancer tissues and the level of ERRα expression was determined to be significantly correlated with the expression level of human epidermal growth factor receptor 2 (HER2). Overexpression of ERRα promoted the proliferation and metastasis of breast cancer in vitro and in vivo while knockdown of ERRα suppressed. Moreover, ERRα was upregulated in chemoresistant breast cancer cells, while ERRα knockdown sensitized cisplatin treatment. Our results further demonstrated that ERRα enhanced the pluripotency of cancer cells and facilitated chemoresistance via upregulating CCNE2 by binding its promoter region. Overexpression of CCNE2 could reverse the sensitivity of breast cancer cells to cisplatin caused by ERRα depletion, thus resulting in accelerated tumor growth. Conclusion We identify that ERRα sustains the chemoresistance of breast cancer cells via regulating the transcription of CCNE2 and may serve as a therapeutic target in breast cancer.

The E3 ubiquitin ligase is an important regulator of cell signaling and proteostasis and is tightly controlled in many diseases, including cancer. Our study aimed to investigate the biological role of the E3 ubiquitin ligase CBLC in breast cancer and elucidate the specific mechanistic network underlying CBLC-mediated target substrate degradation, cell proliferation and metastasis. Here, we showed that CBLC expression was higher in breast cancer tissues and cells than that in normal tissues and cells. Higher expression of CBLC predicted a better prognosis for breast cancer patients. CBLC inhibited the proliferation, migration and invasion of breast cancer cells. Co-IP and immunofluorescence co-localization assays demonstrated that CBLC interacted with CTTN in the cytoplasm. CBLC promoted the degradation of CTTN through the ubiquitin-proteasome pathway without affecting its mRNA level. The inhibitory effect of CBLC on breast cancer cell proliferation, migration and invasion could partly be reversed by CTTN. Taken together, our study clarified the biological role of CBLC as a tumor suppressor and discovered its functional substrate, providing a molecular basis for CBLC/CTTN as a potential therapeutic target in breast cancer.

Background Glutamate metabotropic receptors (GRM) play a variety of roles in neuronal cells. However, their clinical significance and biological functions in breast cancer remain unknown. Methods RNA sequencing data of breast cancer was obtained from the TCGA dataset (v2) and mined for the expression profiles of GRM family according to cancer subtypes. mRNA expression of GRM family in breast cancer tissues and para-cancerous tissue samples as well as breast cancer cell lines were measured by qPCR. The effects of over- and under-expression of GRM4 on cell capabilities to survive, migrate and invade were determined by colony formation, transwell migration and invasion assays. To explore the upstream regulation pattern of GRM4, miRNAs that target GRM4 were predicted and validated by dual luciferase reporter assay. In addition, the mRNA and protein expression of GRM4 regulated by these miRNAs were further measured by qPCR and western blot assay. Results GRM4 was the only GRM member that expressed in breast cancer tissues. Ectopic expression of GRM4 was correlated with better prognosis of breast cancer patients. Overexpression of GRM4 could significantly inhibit cell proliferation, migration and invasion capacity in MDA-MB-231, while knockdown of GRM4 could promote these processes. miR-328-3p and miR-370-3p were predicted to regulate the expression of GRM4 and dual luciferase reporter assay demonstrated that miR-328-3p and miR-370-3p directly bound to the 3′ UTR of GRM4 and mutations on the binding regions on GRM4 significantly decreased the luciferase activity. qPCR demonstrated that expression of miR-328-3p and miR-370-3p was significantly decreased in breast cancer tissues and cells compared with that in control samples. However, there were no correlations between the expression of miR-328-3p and GRM4, as well as the expression of miR-370-3p and GRM4. Moreover, overexpression of miR-328-3p and miR-370-3p counteracted the inhibitory effect of GRM4-induced cell proliferation, migration and invasion. Conclusions Our results suggest that GRM4 might be a tumor suppressor gene in breast cancer under the direct regulation of miR-328-3p and miR-370-3p.

The current protein interaction method is time consuming and cumbersome or the instrument is expensive. A new method that is convenient, fast, and high throughput needs to be studied urgently. The purpose of this study was to establish a homogeneous immunoassay to detect the interaction between insulin-like growth factor-1 receptor-β (IGF1R-β) and suppressor of cytokine signaling 1 (SOCS1). The recombinant vectors IGF1R-β/pENTER and SOCS1/pENTER were constructed and transfected into 293T cells. Based on homogeneous immunoassay technology, we established a suitable method. The signal intensity in the 293T lysate that overexpressed IGF1R-β and SOCS1, respectively, was compared with the signal intensity in the simultaneous expression of IGF1R-β and SOCS1. The interaction between IGF1R-β and SOCS1 was verified in vitro. The detection system for the interaction between IGF1R-β and SOCS1 was established. Compared with other methods, homogeneous immunoassay has the advantages of being rapid and sensitive, having higher sensitivity, and easy to operate. The interaction between IGF1R-β and SOCS1 was tested to verify the feasibility of this method and prove its practicability and sensitivity. This new method can be used as a high-throughput platform for protein-protein interaction, with the advantages of trace detection, short detective time, and high detective sensitivity.

ObjectiveTo investigate the performance of serological test and dynamics of serum antibody with the progress of SARS-CoV-2 infections.MethodsA total of 419 patients were enrolled including 19 confirmed cases and 400 patients from fever clinics. Their serial serum samples collected during the hospitalization were menstruated for IgM and IgG against SARS-CoV-2 using gold immunochromatographic assay and chemiluminescence immunoassay. We investigated whether thermal inactivation could affect the results of antibody detection. The dynamics of antibodies with the disease progress and false positive factors for antibody testing were also analyzed.ResultsThe positive rate of IgG detection was 91.67% and 83.33% using two CLIA, respectively. However, the IgM positive rate was dramatically declined might due to the lack of blood samples at early stages of the disease. The chemiluminescence immunoassay had a favorable but narrow linear range. Our work showed increased IgG values in serums from virus-negative patients and four negative samples were IgG weak-positive after thermal incubation. Our data showed the specificity of viral N+S proteins was higher than single antigen. Unlike generally thought that IgM appeared earlier than IgG, there is no certain chronological order of IgM and IgG seroconversion in COVID-19 patients. It was difficult to detect antibodies in asymptomatic patients suggesting that their low viral loads were not enough to cause immune response. Analysis of common interferent in three IgG false-positive patients, such as rheumatoid factor, proved that false positives were not caused by these interfering substances and antigenic cross-reaction.ConclusionsViral serological test is an effective means for SARS-CoV-2 infect detection using both chemiluminescence immunoassay and gold immunochromatographic assay. Chemiluminescence immunoassay against multi-antigens has obvious advantages but still need improve in reducing false positives.

Orf virus has been utilized as a safe and efficient viral vector against not only diverse infectious diseases, but also against tumors. However, the nature of the genes triggered by the vector in human cells is poorly characterized. Using RNA sequencing technology, we compared specific changes in the transcriptomic profiles in human foreskin fibroblast cells following infection by the orf virus. The results indicated that orf virus upregulates or downregulates expression of a variety of genes, including genes involved in antiviral immune response, apoptosis, cell cycle and a series of signaling pathways, such as the IFN and p53-signaling pathways. The orf virus stimulates or inhibits immune gene expression such as chemokines, chemokine receptors, cytokines, cytokine receptors, and molecules involved in antigen uptake and processing after infection. Expression of pro-apoptotic genes increased at 8 hours post-infection. The p53 signaling pathway was activated to induce apoptosis at the same time. However, the cell cycle program was promoted after infection, which may be due to the immunomodulatory genes of the orf virus. This presents the first description of transcription profile changes in human foreskin fibroblast cells after orf virus infection and provides an in-depth analysis of the interaction between the host and orf virus. These data offer new insights into the understanding of the mechanisms of infection by orf virus and identify potential targets for future studies.

Background: COVID-19 Emerged as a novel zoonotic disease in late 2019 and quickly spread across Wuhan before spreading to other parts of China and rest of the world. Due to the rapid spread of the disease, local hospitals were inundated with COVID-19 patients putting a strain on the healthcare system. Little was known about the transmission and potential clinical management of COVID-19 at that time. Methods: A temporary COVID-19 hospital was built within one week. The confirmed COVID-19 cases were either directly recruited to the hospital or were transferred from other hospitals. Patients were admitted for both quarantine and treatment, as required. Data were collected as part of standard clinical care and retrospectively analyzed. Findings: A total of 2,959 patients were recruited during the operation period of this hospital between February 4, 2020, and April 8, 2020. These patients included 838 severe patients of which 72 were classified as critical, and 66 patients died. No infection was reported among healthcare workers. Interpretation: Setting up a dedicated hospital for COVID-19 provided a critical resource during the peak of the pandemic in Wuhan by enabling both quarantine and treatment for the infected patients. The mortality in this hospital was comparable to other hospitals at the time. These data suggest that this approach may prove beneficial in controlling infectious disease spread and limit mortality and prevent strain on existing healthcare system to enable them to care for non-COVID-19 patients. Funding Statement: This study was supported by funding from Beijing Nova Program Interdisciplinary Cooperation Project (DC; No. Z191100001119021), Chinese PLA General Hospital Youth Project (DC; No.QNF19074), Beijing Nova Program Project (DC; No. Z171100001117012), and China 13th Five-year National Key Grant (LXX; No.2018ZX09201013). Declaration of Interests: The authors declare that there are no competing interests. Ethics Approval Statement: This study was approved by the ethics committee of the Chinese PLA General Hospital, with a waiver of informed consent.

The sodium hydrogen exchanger 1 (NHE1), which functions in maintaining the ratio of Na(+) and H(+) ions, is widely distributed in cell plasma membranes. It plays a prominent role in pH balancing, cell proliferation, differentiation, adhesion, and migration. However, its exact subcellular location and biological functions in Toxoplasma gondii are largely unclear. In this study, we cloned the C-terminal sequence of T. gondii NHE1 (TgNHE1) incorporating the C-terminal peptide of NHE1 (C-NHE1) into the pGEX4T-1 expression plasmid. The peptide sequence was predicted to have good antigenicity based on the information obtained from an immune epitope database. After induction of heterologous gene expression with isopropyl-b-D-thiogalactoside, the recombinant C-NHE1 protein successfully expressed in a soluble form was purified by glutathione sepharose beads as an immunogen for production of a rabbit polyclonal antiserum. The specificity of this antiserum was confirmed by western blotting and immunofluorescence. The antiserum could reduce T. gondii invasion into host cells, indicated by the decreased TgNHE1 expression in T. gondii parasites that were pre-incubated with antiserum in the process of cell entry. Furthermore, the antiserum reduced the virulence of T. gondii parasites to host cells in vitro, possibly by blocking the release of Ca(2+). In this regard, this antiserum has potential to be a valuable tool for further studies of TgNHE1.

Glutamate Ionotropic Receptor Kainate Type Subunit 3 (GRIK3) is an important excitatory neurotransmitter receptor that plays a significant role in various neurodegenerative diseases. However, the biological functions of GRIK3 in malignancies are largely unknown because of limited related studies. Here, we primarily reported that the expression of GRIK3 was higher in breast cancer tissues than in adjacent noncancerous tissues. GRIK3 expression was also positively correlated with the prognosis of patients with breast cancer. GRIK3 promoted the proliferation and migration abilities of breast cancer cells and enhanced the growth of orthotopically implanted tumors. Mechanically, GRIK3 influenced a range of signaling pathways and key signal transducers, including two epithelial‐mesenchymal transition regulators, SPDEF and CDH1. Heterogenous expression of SPDEF and CDH1 counteracted the migration and invasion abilities, respectively, of breast cancer cells induced by GRIK3. Moreover, overexpression of GRIK3 increased the expression of mesenchymal markers and decreased the expression of epithelial markers, resulting in the translocation of β‐catenin into the nucleus and the increased β‐catenin transcriptional activity. In conclusion, the present study reported a novel oncogenic role of GRIK3. Meanwhile, GRIK3, as a membrane receptor, may also serve as a potential therapeutic target for the treatment of breast cancer.

Background Sorafenib is the only approved first line systemic therapy for advanced hepatocellular carcinoma (HCC) in the last decade. Tumour resistance to sorafenib has been of major obstacles to improve HCC patient survival. Methods We polarised THP-1 cells to M1 and M2 macrophages, performed various in vitro assays and developed sorafenib-resistant xenograft models to investigate the role of tumour-associated macrophages (TAM)-secreted molecules in HCC resistance to the targeted therapy. Results We demonstrated M2, but not M1, macrophages not only promote proliferation, colony formation and migration of hepatoma cells but also significantly confer tumour resistance to sorafenib via sustaining tumour growth and metastasis by secreting hepatocyte growth factor (HGF). HGF activates HGF/c-Met, ERK1/2/MAPK and PI3K/AKT pathways in tumour cells. Tumour-associated M2 macrophages were accumulated in sorafenib-resistance tumours more than in sorafenib-sensitive tumours in vivo and produced abundant HGF. HGF chemoattracts more macrophages migrated from surrounding area, regulates the distribution of M2 macrophages and increases hepatoma resistance to sorafenib in a feed-forward manner. Conclusions Our results provide new insights into the mechanisms of sorafenib resistance in HCC and rationale for developing new trials by combining sorafenib with a potent HGF inhibitor such as cabozantinib to improve the first line systemic therapeutic efficacy.

Few prognostic indicators with differential expression have been reported among the differing ER statuses. We aimed to screen important breast cancer prognostic genes related to ER status and to construct an efficient prognostic prediction system. mRNA expression profiles were downloaded from TCGA and GSE70947 dataset. Two hundred seventy-one overlapping differentially expressed genes (DEGs) between the ER- and ER+ breast cancer samples were identified. Among the 271 DEGs, 109 prognostically relevant mRNAs were screened. mRNAs such as RASEF, ITM2C, CPEB2, ESR1, ANXA9, and VASN correlated strongly with breast cancer prognosis. Three modules, which contained 28, 9 and 8 enriched DEGs, were obtained from the network, and the DEGs in these modules were enriched in response to hormone stimulus, epithelial cell development, and host cell entry. Using bayes discriminant analysis, 48 signature genes were screened. We constructed a prognostic prediction system using the 48 signature genes and validated this system as relatively accurate and reliable. The DEGs might be closely associated with the prognosis in patients with breast cancer. We validated the effectiveness of our prognostic prediction system by GEO database. Therefore, this system might be a useful tool for preliminary screening and validation of potential prognosis indicators for ER+ breast cancer derived from mechanistic research.

Background The Orf virus (ORFV) is the causative agent of orf, a globally-occurring, acute, pustular, contagious disease affecting sheep, goats and humans with a worldwide distribution. Currently, the genomic analysis of four ORFV strains from the Fujian province in southern China and a NA1/11 strain isolated from the Jilin province in northeast China have been reported. However, little is known about the genomic information of ORFV strains from central China. Results From a recent outbreak in a sheep herd in the Henan province of central China, a novel ORFV strain (HN3/12) was isolated and cultured in ovine fetal turbinate (OFTu) cells. The strain was identified as HN3/12 and verified by PCR based on the DNA sequences of 011 and 059 genes. The whole genomic sequence of this isolate was determined by Next Generation Sequencing technology. To determine the genetic characteristics of the HN3/12 strain, phylogenetic analysis of the 011 and 059 genes and amino acid sequence alignment of the HN3/12 strain were performed and compared with reference parapoxvirus strains. Conclusions The HN3/12 genome is 136,643 bp in length, contains 63.67% G + C and encodes 132 putative genes. Phylogenetic analysis of the 011 and 059 nucleotide sequences showed that this viral strain was similar to the NA1/11 isolate. The homology analysis indicates that HN3/12 has 93% to 98% identity with published ORFV strains at amino acid level. When open reading frames (ORFs) were aligned among the HN3/12 and four Fujian ORFV strains, most of them have identities greater than 90% and only a few less than 60%. The availability of the whole genomic sequence of HN3/12 aids in our understanding of, and provides new insights into, the genetic diversity of ORFV. Electronic supplementary material The online version of this article (doi:10.1186/s12917-017-1178-1) contains supplementary material, which is available to authorized users.

SRY-related high-mobility-group box 9 (SOX9) is a member of the SOX family of transcription factors. Accumulating evidence has shown that SOX9 plays a significant role in various malignancies. However, the role of SOX9 in nasopharyngeal carcinoma (NPC) remains unknown. In the present study, up-regulation of SOX9 was observed in both NPC tissues and different NPC cells. Overexpression of SOX9 promoted NPC cell proliferation, migration and invasion. Conversely, knock down of SOX9 inhibited NPC proliferation, colony formation, migration and invasion. Mechanistically, SOX9 bound directly to the promoter region of BMP2 and increased BMP2 expression. In addition, overexpression of SOX9 activated the mTOR pathway partly through BMP2. Collectively, these results identify a novel role for SOX9 as a potential therapeutic marker for the prevention and treatment of NPC.