Expression And Biological Significance Of G Protein Coupled Receptor 34 In Patient With Non-small Cell Lung Cancer

Objective: This study is to investigate the use of bioinformatic methods to analyze the differential expression of GPR34 gene and protein in normal lung tissue and non-small cell lung cancer(NSCLC)tissue,to analyze the relationship between GPR34 and NSCLC prognosis;This study is to investigate GPR34 gene expression by biological experiments,to analyze the correlation of GPR34 and NSCLC;This study is to stimulate the GPR34 gene expression by gene overexpression to investigate the changes in cell growth,invasion and migration.Methods:1?Combined with analysis of Oncomine,TCGA and HPA,the expression levels of GPR34 gene and protein in normal and NSCLC tissues were systematically investigated.In addition,we also analyzed the correlation of GPR34 gene expression and NSCLC prognosis by the survival model of Kaplan-Meier database.2?The expression of GPR34 mRNA was examined by qRT-PCR in 70 paired NSCLC tumor tissues and its matched adjacent normal tissues.We compared the differential expression between them,and further analyzed the correlation of GPR34 and clinicopathologic characteristics.3 ? The expression of GPR34 and Ki67 protein were examined by immunohistochemistry in NSCLC tumor tissues of 70 cases and adjacent normal lung tissues of 70 cases;We analyzed the correlation of GPR34,Ki67 and clinicpathologic characteristics respectively,and further discussed the relationship between GPR34 and Ki67.4?The expression of GPR34 gene and protein in a variety of NSCLC cell lines was detected by qRT-PCR and Western blot.5?The GPR34 overexpression vector was designed and transfected into lung adenocarcinoma A549 cell line.The expression of GPR34 mRNA was detected by qRT-PCR.Cell proliferation was detected by CCK8.Cell invasion and migration were detected by Transwell.Results:1?GPR34 gene and protein were found to be down-regulated in NSCLC by in silico analysis,and its low-expression correlated with poor prognosis in patients with NSCLC,especially lung adenocarcinoma.2?The expression of GPR34 mRNA in NSCLC tissues were 0.46+0.24 times lower than that in normal lung tissues.The expression of GPR34 was related with tumor differentiation of NSCLC.3?GPR34 protein was found to be down-regulated in NSCLC tissues whereas up-regulated in normal lung tissues.Ki67 protein was found to be up-regulated in NSCLC tissues whereas down-regulated in normal lung tissues.The expression of GPR34 was related with tumor differentiation.Ki67 expression was correlated with tumor differentiation.A certain degree of negative correlation exists between GPR34 and Ki67 expression in NSCLC tissues.4?Combined with the results of qRT-PCR and Western blot,GPR34 gene and protein were found to be down-regulated in several NSCLC cell lines.5?After transfection of GPR34 overexpression vector into lung adenocarcinoma A549 cell line,GPR34 gene expression was significantly up-regulated as detected by qRT-PCR.The overexpression of GPR34 gene decreases proliferation,invasion and migration ability of A549 cell.Conclusion:1?The expression of GPR34 gene in NSCLC was analyzed by bioinformatics metheds,and the results indicated that GPR34 gene was down-regulated in NSCLC.2?It was confirmed that GPR34 was the NSCLC-related gene for the first time,and its down-regulated expression was closely related to the NSCLC differentiation and poor prognosis3?GPR34-specific overexpression vector may efficiently up-regulate the expression of GPR34 gene after transfected into A549 cell.GPR34 gene overexpression inhibits the proliferation,invasion and migration ability of A549 cell.