Preliminary Study On The Function And Mechanism Of Liver Cancer-related Genes FBXW8 And RTEL1

Part 1: The effects of FBXW8 on cells proliferation and migration of hepatocellular carcinoma cell linesObjective: To investigate the effects of F-box and WD repeat domain containing 8(FBXW8)on the proliferation and migration of hepatocellular carcinoma(HCC)cell lines HepG2 and Huh7 and the underlying molecular mechanism.Methods: The FBXW8 overexpression recombinant plasmids were transfected into the HepG2 and Huh7 cells.The small interfering RNAs(siRNAs)targeting FBXW8 were used to knockdown the endogenous FBXW8 in HepG2 and Huh7 cells.Western blotting(WB)assays were used to detect the protein levels of FBXW8 in HepG2 and Huh7 cells to verify the effects of its overexpression or knockdown;qRT-PCR assays were used to detect the protein expression levels of several markers relevant to epithelial-mesenchymal transition(EMT)in HepG2 and Huh7 cells.Cell counting kit-8(CCK-8)assays were used to assess the HepG2 and Huh7 cells proliferation;and the transwell assays were used to assess the HCC cells migration.Log-rank test was performed to assess the difference of survival rates between the HCC patients with high and low expression levels of FBXW8.Results: Overexpression of FBXW8 significantly reduced the proliferation and migration abilities of HepG2 and Huh7 cells;whereas knockdown of FBXW8 significantly induced their abilities of proliferation and migration.Overexpression of FBXW8 significantly reduced EMT of HepG2 and Huh7 cells,whereas knockdown of FBXW8 promoted their EMT.FBXW8 was down-regulated in HCC tissues compared to thematched non-tumor liver tissues,and the lower expression levels of FBXW8 predicts poor clinical outcomes of HCC patients.Conclusions: FBXW8 might function as a tumor suppressor in the development of HCC by reducing the HCC cells proliferation,EMT and migration.Part 2: The effects of RTEL1 on cells proliferation and migration of hepatocellular carcinoma cell linesObjective: To investigate the effects of regulator of telomere length helicase 1(RTEL1)on the proliferation and migration of hepatocellular carcinoma(HCC)cell lines HepG2 and Huh7,and the underlying molecular mechanism.Methods: The RTEL1 overexpression recombinant plasmids were transfected into the HepG2 and Huh7 cells.The small interfering RNAs(siRNAs)targeting RTEL1 were used to knockdown the endogenous RTEL1 in HepG2 and Huh7 cells.Western blotting(WB)assays were used to detect the protein levels of RTEL1 in HepG2 and Huh7 cells to verify the effects of its overexpression or knockdown.Cell counting kit-8(CCK-8)assays were used to assess the HepG2 and Huh7 cells proliferation;and the transwell assays were used to assess the HCC cells migration.Results: Overexpression of RTEL1 significantly promoted the proliferation and migration in HepG2 and Huh7 cells;whereas knockdown of RTEL1 significantly reduced their proliferation and migration.Conclusions: RTEL1 might function as a oncogene in the development of HCC by promting the HCC cells proliferation and migration.