The Correlation Between The Expression Of MGMT And Sensitivity To Oxaliplatin In Hepatocellular Carcinoma Cell

Objective:To explore the relationship between the expression of MGMT in hepatocellular carcinoma cell and sensitivity to oxaliplatin.Methods: Bel-7402, SMMC-7721 and HUH-7 were cultured in vitro. RT-PCR was used to detect the MGMT mRNA expression levels, the expression level of MGMT protein was detected by Western-Blot method, Cell apoptosis was detected by flow cytometry and the expression of r-H2 Ax was detected With the method of Western-Blot to evaluate DNA damage and degree after oxaliplatin treatment, the MGMT methylation status of all cells was detected by methylation specific PCR in contrast to the hepatoma cell line MTT.Results:The hepatocellular carcinoma cell line could be detected in the MGMT gene and protein expression, which Huh-7 MGMT mRNA and protein level was the highest; and BEL-7402 MGMT mRNA and protein levels were lowest, and that of the three MGMT protein and mRNA level was positively. The expression of r-H2 Ax was different with oxaliplatin 12 h, which Bel-7402 r-H2 Ax increased significantly while HuH-7 increased at least. According to MTT results to calculate the IC50 of Huh-7, SMMC-7721 and BEL-7402 246.76 u g / ml and 48.43 u g / ml and 31.35 mu g / ml respectively. According to the correlation analysis that MGMT mRNA and protein expression levels and IC50 was positively correlated, the higher MGMT expression level was, the more value IC50 was. Apoptosis was detected by flow cytometry, the early apoptosis rate after adding oxaliplatin showed the highest Bel-7402, and the apoptosis rate of HuH-7 cells was the lowest. MGMT methylation results showed the expression of Huh-7 MGMT methylation level was the lowest, BEL-7402 MGMT methylation expression level is the highest, according to the correlation analysis of MGMT promoter methylation level and Mgmt mRNA and protein levels and IC50 was negatively correlated.Conclusion:The status of MGMT gene may affect the sensitivity of HCC cells to oxaliplatin.The lower of MGMT expression in the hepatocellular carcinoma cell mRNA and protein expression level, the stronger damage of oxaliplatin. The higher the methylation level of MGMT promoter in hepatocellular carcinoma cells, the more sensitive was oxaliplatin; the lower level of the MGMT promoter methylation was, the less sensitive oxaliplatin was.