Mechanism Of Rbm14-enhanced P23 Transcriptional Regulation Of CXCL1-induced EMT In Lung Cancer

Background: Lung cancer is one of the most diagnosed cancers worldwide and the leading cause of cancer-related death.In recent years,lung cancer has become the largest cancer in China,and the onset of cancer shows a trend of young people.The most important reason is its distant metastasis,which is also the main reason for the low overall survival of lung cancer and the poor quality of life of patients.EpithelialMesenchymal transition(EMT)is characterized by loss of epithelial cell adhesion and acquisition ability of migration and invasion of mesenchymal cell.Numerous studies have shown that EMT can coordinate the interaction between multiple characteristics of tumors,such as stem tumor cells,tumorigenicity,resistance to treatment,and adaptability to changes in the microenvironment.Studies have shown that tumor cells that have undergone EMT are more aggressive,bringing more challenges to clinical treatment and the quality of life of cancer patients.Prostaglandin E synthase 3(PTGES3,also known as p23)has been known and studied as a chaperone protein,and it has been reported that p23 is overexpressed in lung adenocarcinoma,and its overexpression degree is positively correlated with the reduction in overall survival of lung adenocarcinoma patients.Our previous studies have confirmed that p23 can be used as a transcriptional factor to participate in the regulation of tumor-related inflammatory processes in lung adenocarcinoma tumor,and provide a theoretical basis for the diversification of p23 functions.In this study,we explore the effect and mechanism of p23 on lung cancer metastasis from the perspective of EMT.Methods: In this research human lung cancer cell lines A549 and H1299 were used as models in vitro.Lentivirus infection was used to construct H1299 cell lines with stable transmutation expressing p23 and A549 cell lines with stable transmutation low p23.The effect of p23 on the EMT phenotype of lung cancer cells was detected by real-time label-free migration detection methods,transwell,scratches and other methods.The effect of p23 on the expression of EMT signature proteins(E-Cadherin,N-Cadherin and Vimentin)was detected by q PCR and Western blot.The potential downstream target gene of p23,CXCL1,was screened by RNA＿sequencing.In the study of molecular mechanism,the transcriptional regulation and potential mechanism of p23 on CXCL1 were detected by DNA-protein binding pulldown analysis method,dual luciferase reporter,gel migration and other experiments.The key protein Rbm14,which interacts with p23,was identified by co-immunoprecipitation combined with mass spectrometry,and its role in p23 regulation of CXCL1-mediated EMT was explored.Results: p23 is overexpressed in clinical lung cancer tissues and lung cancer cell lines,especially in tissues where metastasis occurs.Knockdown p23 significantly inhibits the EMT phenotype of lung cancer cell lines,and overexpression of p23 significantly promotes EMT.The downstream target gene of p23,CXCL1,was screened and identified by RNA＿sequencing,and the p23-mediated EMT process is CXCL1-dependent.The protein that interacts with p23,Rbm14,was identified by mass spectrometry,which can promote the transcriptional regulation of p23 on CXCL1 and thus promote EMT.Conclusion: By forming a protein complex with p23,Rbm14 promotes p23 to regulate the expression of CXCL1,thus promotes the occurrence and development of EMT.Our research shows that the Rbm14-p23-CXCL1-EMT axis has potential application value in the treatment of tumor metastasis(such as lung cancer metastasis).