| Obejective To observe the expression of TSC2in Human hepatocellularcarcinoma HepG2cells and investigate the mechanism of the expression.Methods1. After treatment with different concentration of5-Aza-CdR(0,0.4,1.6,6.4,25.6and102.4μmol/L), the growth inhibitory rate of HepG2cellswas assessed by MTT assay(24h,48h,72h).2. PT-PCR and Western blotting wereapplied to measure the expression of TSC2mRNA and tuberin protein inhepatocellular carcinoma HepG2cells that were treated by5-Aza-CdR.3.Methylationspecific PCR were used to detect respectively the methylation status of TSC2gene intwo groups of HepG2cells treated by medium and5-Aza-CdR.Results1.MTT assay demonstrated that liver cancer HepG2cells proliferationwere inhibited obviousily by5-Aza-CdR. After HepG2cells were treated by diferent5-Aza-CdR concentration (0.4,1.6,6.4,25.6and102.4μmol/L)72hours, theinhibition rates were (40.14±2.11)%,(48.72±3.21)%,(56.66±2.43)%,(62.24±2.43)%and (70.10±3.12)%respectively. While the same concentration5-Aza-CdR of102.4μmol/L, with processing time, the liver cancer HepG2cells proliferationinhibition rates were （62.90±2.14）%,（67.52±2.05）%and（70.10±3.12）%respectively(P <0.05).2.Different concentration of5-Aza-CdR role in HepG2cells,the result showed that after72hours, with the increasing of the concentration,expression of TSC2mRNA and tuberin protein all increased (P<0.05).3. By MSP,TSC2gene of hepatocellular carcinoma HepG2cells treated by medium methylationand demethylation coexist, and methylation level is higher than demethylation, but themethylation level was decreaming in5-Aza-CdR treated group.Conclusions TSC2, as a kind of tumor suppressor gene, decreased in HepG2cells. Its mechanism might be due to that TSC2gene promoter methylation status. |
