Abstract
Objective To observe effects of over-expression of melanoma antigen gene (MAGE) A3 on the progression,apoptosis,metastasis and invasion in human hepatocellular carcinoma cell lines,and investigate its effect on hepatocellular carcinoma (HCC) progession.Methods The lentivirus vector with recombinant MAGE-A3 gene (LV-MAGE-A3) were constructed and transfected into HepG2 and MHCC-97H cell lines and cell only or transfected with empty vector were as controls.The expression of green fluorescent protein was observed by inverted fluorescent microscope to detect tranduced efficiency.MAGE-A3 gene and protein expression was assayed by reverse transcription-polymerase chain reaction (RT-PCR) and Western-Blot technology.The effects of MAGE-A3 over-expression on proliferation,migration and invasion of HCC were detected by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays,clone formation assays and transwell technique.Cell apoptosis was detected by terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL) and cell cycle was analysed by flow cytometry (FCM).Results After lentivirus vector transfection for 120 hours,the expression of green fluorescent protein was the strongest and most stable in cells.The transfection efficiency was up to 80％.After transfected with LV-MAGE-A3,the expression of MAGE-A3 gene and the protein in HepG2 and MHCC-97 cells was higher than that of controls.The cell numbers of migration and invasion were 187.00 ± 13.52,121.00 ±4.16 in HepG2 and 164.00 ±9.64,59.50 ± 1.00 in MHCC-97,which significantly increased compared to control group (P ＜0.01).The cell numbers of S stage increased by FCM.However,the cell proliferation,colony-forming ability and apoptosis of HCC cell lines had no statistical significance between LV-MAGE-A3 group and control (P ＞ 0.05).Conclusion MAGE-A3 gene promote the abilities of migration and invasion in HCC cell lines.It suggests that MAGE-A3 expression is closely corelated to HCC progression. Key words: Melanoma antigen gene ; Hepatocelluar carcinoma; Migration; Invasion
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