Effect Of Hypoxia On The Growth, Migration And Invasion Of Hepatocellular Carcinoma Cells

Objective:To investigate whether the hepatocarcinoma cells cultured under hypoxic conditions have obvious changes in proliferation,apoptosis,migration and invasion ability,and epithelial-mesenchymal transition.Methods:Two kinds of MHCC97 H and Hep G2 hepatocarcinoma cells with different metastasis capacity were selected.The hypoxia model was established by hypoxia culture.The expression of HIF-1? m RNA under normoxia and hypoxia was detected by real-time fluorescent quantitative PCR.The cells were observed under inverted microscope.Morphological changes;MTT assay was used to detect the changes of cell proliferation,flow cytometry was used to detect the changes of apoptosis rate in two groups,scratch test was used to detect the changes of cell migration ability,and Matrigel coated Transwell chamber was used to detect two groups of cells.The invasive ability of the cells was detected by Western Blot assay.The expression of HIF-1?,E-cadherin and Vimentin protein in the two groups was detected.Results:The expression of HIF-1? m RNA in the two groups of MHCC97 H and HepG2 hepatoma cells was significantly higher at 48 h after hypoxia by real-time fluorescence quantitative PCR(P<0.001);the cells in the two groups were changed from tightly packed to more loose cells after 48 h hypoxia.Spindle-like changes were observed,and cells were loose;MTT assays showed greater cell proliferation after hypoxia(P<0.01);flow cytometry suggested that the rate of apoptotic cells in both groups was lower than that in normoxia group(P <0.05);Scratch test showed that after the two groups of hepatoma cells hypoxia,their migration ability was significantly increased(P<0.05);Transwell experiments showed that after the two groups of hepatoma cells hypoxia,their invasion ability was significantly enhanced(P <0.05);Western Blot showed that the two groups of cells were HIF-1? protein.The expression of E-cadherin in epithelial cells was significantly decreased after hypoxia(P<0.01),and the expression of Vimentin was significantly increased(P<0.05).Conclusion:Under certain hypoxic conditions,the proliferation ability of hepatoma cells was increased,the apoptosis rate was decreased,and the migration and invasiveness were enhanced,and epithelial-mesenchymal transition occurred.