MicroRNA Hsa-let-7c-5p Via Targeting TK1 Affects The Proliferation,Migration And Invasion Of Hepatocellular Carcinoma

Objective: To explore the mechanism of microRNA(microRNA)hsa-let-7c-5p on the proliferation,migration and invasion of hepatoma cells by regulating targeted thymidine kinase 1(TK1).Methods:The GEPIA database was used to analyze the expression difference and prognosis of TK1 in liver cancer.The mRNA expression of TK1 in liver cancer tissues,adjacent normal tissues,human normal hepatocyte LO2 and human hepatoma cell lines was detected by real-time quantitative PCR(qRT-PCR).The expression of TK1 protein in human normal hepatocyte LO2 and human hepatoma cell lines was detected by Western Blot.Human hepatoma cells HUH-7 and MHCC-97H were selected as the main objects of this study.By transfecting TK1 overexpressing and knockdown lentiviruses.The transfection efficiency was detected by qRT-PCR and Western Blot experiments.The effects of TK1 on the proliferation of HUH-7 and MHCC-97H cells were detected by CCK-8 experiments.The effect of TK1 on the migration and invasion of HUH-7 and MHCC-97H cells was detected by Transwell experiments.The effects of TK1 on the expression levels of epithelial-mesenchymal transition-related proteins and cell cycle-related proteins in HUH-7 and MHCC-97H cells were detected by Western Blot.The potential regulatory relationship between hsa-let-7c-5p and TK1 was predicted by the starbase website,and the binding relationship between hsa-let-7c-5p and TK1 was verified by dual-luciferase reporter gene experiments.The expression of hsa-let-7c-5p in liver cancer tissues,adjacent normal tissues,human normal hepatocyte LO2 and human hepatoma cell lines was detected by real-time quantitative PCR(qRT-PCR).The hsa-let-7c-5p mimics and inhibitor were transfected into MHCC-97H and HUH-7 cells.The transfection efficiency was detected by real-time quantitative PCR(qRT-PCR).The effect of hsalet-7c-5p on the proliferation of HUH-7 and MHCC-97H cells was detected by CCK-8 experiments.The effect of hsa-let-7c-5p on the migration and invasion of HUH-7and MHCC-97H cells was detected by Transwell assay.The effects of hsa-let-7c-5p on the expression levels of epithelial-mesenchymal transition-related proteins and cell cycle-related proteins in HUH-7 and MHCC-97H cells were detected by Western Blot.TK1 lentivirus,hsa-let-7c-5p mimics and hsa-let-7c-5p inhibitor were cotransfected into HUH-7 and MHCC-97H cells.TK1 overexpressing and knockdown lentivirus,hsa-let-7c-5p mimics and hsa-let-7c-5p inhibitor were co-transfected into HUH-7 and MHCC-97H cells.The expression changes of TK1 in HUH-7 and MHCC-97H cells were detected by real-time quantitative PCR(qRT-PCR)and Western Blot.The changes of the proliferation of HUH-7 and MHCC-97H cells after co-transfection with lentivirus and mimics or inhibitor were detected by CCK-8experiments.The changes of migration and invasion of HUH-7 and MHCC-97H cells after co-transfection of lentivirus and mimics or inhibitor were detected by Transwell assay.Changes in the expression levels of epithelial-mesenchymal transition-related proteins and cell cycle-related proteins in HUH-7 and MHCC-97H cells after cotransfection of lentivirus and mimics or inhibitor were detected by Western Blot.Results: The results of the GEPIA database indicated that TK1 was highly expressed in hepatocellular carcinoma and correlated with prognosis.The results of qRT-PCR showed that TK1 was highly expressed in liver cancer tissues and hepatoma cell lines.The results of Western Blot showed that TK1 was highly expressed in hepatoma cell line.The results of CCK-8 experiments showed that TK1 could promote the proliferation of hepatoma cells.Transwell experiments showed that TK1 could promote the migration and invasion of hepatoma cells.Western Blot showed that TK1 could promote the expression of Cyclin A,Cyclin E and E-cadherin,and inhibit the expression of P21 and Vimentin.According to the results of starbase and dualluciferase reporter gene experiments,TK1 is transcriptionally regulated by the microRNA hsa-let-7c-5p.And the expression of hsa-let-7c-5p and TK1 was negatively correlated.The results of GEPIA database showed that hsa-let-7c-5p was lowly expressed in hepatocellular carcinoma and correlated with prognosis.The results of qRT-PCR showed that hsa-let-7c-5p was lowly expressed in liver cancer tissues and hepatoma cell lines.The results of CCK-8 experiments showed that hsalet-7c-5p could inhibit the proliferation of hepatoma cells.The results of Transwell experiment showed that hsa-let-7c-5p could inhibit the migration and invasion of hepatoma cells.The results of Western Blot experiments showed that hsa-let-7c-5p could promote the expression of P21 and Vimentin,and inhibit the expression of Cyclin A,Cyclin E and E-cadherin.After co-transfection of lentivirus and mimics with inhibitor,the results of qRT-PCR and Western Blot showed that hsa-let-7c-5p could inhibit the expression of TK1.The experimental results of CCK-8 showed that hsa-let-7c-5p could affect the proliferation of hepatoma cells by regulating TK1.The results of Transwell experiments showed that hsa-let-7c-5p could affect the migration and invasion of hepatoma cells by regulating TK1.The results of Western Blot experiments showed that hsa-let-7c-5p could affect the effects of Cyclin A,Cyclin E,P21,E-cadherin and Vimentin by regulating TK1.Conclusions:TK1 is highly expressed in liver cancer tissues and hepatoma cell lines,and can promote the proliferation,migration and invasion of hepatoma cell,and can also promote the expression of Cyclin A,Cyclin E and E-cadherin,and inhibit the expression of p21 and Vimentin.Hsa-let-7c-5p is lowly expressed in liver cancer tissues and hepatoma cell lines,and can inhibit the proliferation,migration and invasion of hepatoma cell,promote the expression of p21 and Vimentin,and inhibit expression of Cyclin A,Cyclin E and E-cadherin.The expressions of hsa-let-7c-5p and TK1 were negatively correlated,and hsa-let-7c-5p could target and regulate the expression of TK1.