The Relationship Between Hypoxia And Epithelial Mesenchymal Transition In Human Liver Cancer HepG-2 Cells

Objective: To research the expression of hypoxiainduced factor-1 alpha、 molecular related epithelial mesenchymal transition E-cadherin 、N-cadherin in Hep G-2 cells, and discuss the relationship between hypoxia and epithelial mesenchymal transition, through cobalt chloride chemical manufacturing hypoxia microenvironment.Methods: Using the cobalt chloride chemical induced hypoxia model of Hep G-2 cells, we detect proliferation activity of cell through MTT method and detect HIF-1 alpha, E-cadherin, N-cadherin m RNA and protein expression level through q PCR method and Western blotting method in Hep G-2 cells.Results: MTT showed that the growth inhibition was of varying degrees respectively in different concentration groups of cells and different time groups of cells after Hep G-2 cells were treated with cobalt chloride. The inhibiting effect on the proliferation of different time groups of Hep G-2 cells was the most significant with the use of cobalt chloride of 250μmol/L, the OD value was lower than that of other concentration groups of cells and the proliferation inhibition differences between cobalt chloride of 250μmol/L and the control group, 50μmol/L group, 100μmol/L group, 200μmol/L group and 300μmol/L group were statistically significant(P<0.05). Cobalt chloride of 50μmol/L and 200μmol/L inducing hypoxia, Hep G-2 cells growth were in good condition and there were obvious cell proliferation. When treated with cobalt chloride of 50μmol/L, the OD value of Hep G-2 cells was higher than that of other groups, and the differences in the OD value of Hep G-2 cells treated with cobalt chloride of 50μmol/L, 100μmol/L, 150μmol/L, 250μmol/L and 300μmol/L, all showed statistical significance(P<0.05). When treated with cobalt chloride of 200μmol/L, the OD value of Hep G-2 cells was higher than that of Hep G-2 cells treated with cobalt chloride of 100μmol/L, 150μmol/L, 250μmol/L and 300μmol/L, and the differences all showed statistical significance(P<0.05). q PCR and Westernblotting showed that HIF-1αm RNA was higher than that of the control group, with statistical significant difference(P<0.05), the expression of the experimental group E-cadherin m RNA was lower than that of the control group, with statistical significant difference(P<0.05), the expression of the experimental group N-cadherin m RNA was higher than that of the control group, but the difference was not statistically significant(P=0.41), however, there was a tendency to show statistical difference. The protein expression of the experimental group HIF-1αand N-cadherin m RNA was higher than that of the control group, with statistical significant difference(P<0.05); the protein expression of the experimental group E-cadherin m RNA was lower than that of the control group, with statistical significant difference(P<0.05).Conclusions: 1.Tumor hypoxia microenvironment can be simulated on condition that Hep G-2 cells are treated with cobalt chloride of 200μmol/L for 24 hours.2. q PCR and Western blotting showed that hypoxia can up-regulate the expression of HIF-1αand N-cadherin and down-regulate the expression of E-cadherin.3. Hypoxia can trigger the epithelial mesenchymal transition(EMT) in liver cancer cells.