The Preliminary Study On MiRNA Targeting Gab2 In Hepatocellular Carcinoma Cell

Hepatocellular carcinoma(HCC)is one of the most popular malignancies in the world,and its high mortality rate and high recurrence rate have always been the problems plaguing clinical treatment.With the increased understanding of the molecular mechanisms of HCC development and therapeutic resistance,clinical researchers have developed many molecular drugs.These drugs may target one or more of the key signaling pathways in cancer development,such as cell pro liferation,apoptosis and angiogenesis.The adapter protein Gab2,as a scaffold molecule,not only plays an important role in pathological processes such as abnormal proliferation,invasion and migration of cells,but also plays an important role in signal transduction,which makes it play an important role in the occurrence and development of human cancer.Previous studies in our laboratory showed that the expression of Gab2 in hepatocellular carcinoma was significantly higher than that in paracancerous or normal tissue,and the proliferation,migration and invasion ability of Gab2-overexpressing tumor cells were often higher than that of adjacent paracancerous or normal cells.The high expression of Gab2 in hepatocellular carcinoma cells is mediated by many factors and regulatory mechanisms.However,the regulation of microRNA expression is crucial,and the abnormal expression of microRNA as a biomarker can be used in the early diagnosis and prognosis of HCC.Therefore,we explored the microRNA targeting Gab2.In this study,we used bio informatics website and software to predictel the microRNAs that may regulate Gab2,9 microRNAs were screened and detected their expression in HepG2,HL-7702 and Gab2-related hepatoma cell lines,and further screened four microRNAs related to Gab2 expression.Subsequently,we trancfected miRNA mimic to detect the expression of Gab2 protein.The results showed that Gab2 was the downstream target of hsa-miR-9 and hsa-miR-181a,hsa-miR-9 and hsa-miR-181a could steadily inhibit the expression of Gab2 protein.At the same time,we constructed recombinant plasmids and mutation binding site plasmids containing the predicted binding sites of hsa-miR-9,hsa-miR-181a and Gab2 gene 3’UTR,respectively,and determined their binding sites by dual luciferase reporter system.Then,we detected the expression of Gab2 mRNA by trancfecting hsa-miR-9 mimic and hsa-miR-181a mimic,and explored the molecular mechanism of hsa-miR-9 and hsa-miR-181a targeting Gab2.In addition,we also found that overexpression of hsa-miR-9 and hsa-miR-181a can affect the migration and proliferation of hepatoma cells,suggesting that it may play a role in tumor suppressor by targeting Gab2 in hepatoma cells.It provides a valuable theoretical reference for the relationship between Gab2 and signal transduction and regulation of the progression of liver cancer,and provides a new idea for the prognosis and prognosis of hepatocellular carcinoma.