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Triptolide down-regulates P53 gene methylation and inhibits proliferation of hepatocarcinoma SMMC-7721 cells
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Abstract:
Objective: To study the effect of triptolide (TP) on the proliferation of hepatocarcinoma SMMC-7721 cells and its effect on demethylation of P53 gene. Methods: The effect of TP on proliferation of SMMC-7721 cells was measured by MTT method, the effect of TP on P53 gene methylation in SMMC-7721 cells was analyzed by methylation specific PCR, the expressions of methyltransferase DNMT1, DNMT3a and DNMT3b mRNA in SMMC-7721 cells were measured by RT-PCR, and the protein expression of P53 in SMMC-7721 cells was detected by Western blotting assay. Results: TP inhibited the proliferation of SMMC-7721 cells in a dose-dependent manner (P<0.05), with the inhibitory rate being (73.5±3.02)% at 40 ng/ml TP, and IC50 of TP was 20 ng/ml. TP significantly inhibited DNMT1, DNMT3a, and DNMT3b mRNA expressions in SMMC-7721 cells (P<0.05, P<0.01), and dose-dependently reversed the hypermethylation of P53 gene and enhanced P53 protein expression in SMMC-7721 cells. Conclusion: TP can inhibit the proliferation of SMMC-7721 cells through the inhibiting methyltransferase expression, which augment the demethylation of P53 gene and results in the increased expression of P53 protein.
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Project Supported:
Project supported by the National Natural Science Foundation of China (No. 81072444)
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