The Influence Of ADAMT-18 Expression On The Proliferation And Invasiveness Characteristics On The Hepatocellular Carcinoma Cell Lines MHCC-97H And HepG2

Objective: We chose ADAMTS-18 gene from our previous Genome-wide association study,and to explore mechanism and effects of up or down-regulation of the gene on proliferation and invasion in hepatocellular carcinoma cell lines,MHCC-97 H and HepG2.Methods: We collected 62 pairs of tumor and non-tumor tissue samples of patients in 2016,which was diagnosed with HBV related HCC and received surgical operation by departments of pathology and hepatobiliary of the first hospital affiliated Guangxi medical university.(1)We extracted RNA from tumor and non-tumor tissue by Trizol,and respectively detected the expression of ADAMTS-18 in tissue by qRT-PCR and immunohistochemical method.Then we analysed the correlation between the expression of ADAMTS-18 and clinical pathological characters.(2)We then extracted RNA and detected the expression of the gene through the same methods,and selected cell lines with remarkable expression of ADAMTS-18.(3)Expression plasmid and silence plasmid wererespectively synthesized and transfected into MHCC-97 H,HepG2 and HL-7702 cell lines.And the transfection efficiency was assessed by the fluorescence microscopy,qRT-PCR and Western blot.(4)The changes of proliferation and invasiveness in MHCC-97 H,HepG2 and HL-7702 cell lines were detected by MTT method,Wound healing assay and Transwell chamber in vitro invasion assay.Results: We found the expression of ADAMTS-18 in tumor tissue was significantly higher than that in non-tumor tissue(P <0.05).The expression of ADAMTS-18 mRNA was detected in all six cell lines in the study.We completed to construct stable cell lines(MHCC-97 H,Hep G2 and HL-7702)of over-expression and down-regulated expression for ADAMTS-18.Wound healing assay and Transwell chamber assay showed that significant enhancement of migratory and invasion abilities were found in MHCC-97 H and HepG2 with over-expression of ADAMTS-18(P <0.05).While a remarkable decline of invasion was only found in MHCC-97 H with down-regulated expression for ADAMTS-18(P <0.05).MTT method showed a significant increase of proliferation ability was found in MHCC97H、HepG2 and HL-7702 with over-expression of ADAMTS-18(P <0.05).Conclusion: The expression of ADAMTS-18 in HCC tissue was significantly higher than that in non-tumor tissue,which may promote the proliferation on MHCC97 H,HepG2,HL-7702 cell lines,and promote migratory and invasion abilities in MHCC97 H cell lines.