雷公藤内酯醇逆转人恶性淋巴瘤CA46细胞系p16基因甲基化的实验研究

Experimental Study on Triptolide Reversing Hypermethylation of P16 in Malignant Lymphoma Cell Line CA46

目的:以p16基因存在异常甲基化的恶性淋巴瘤细胞CA46为研究对象,探究雷公藤内酯醇(triptolide,TPL)对P16这一细胞周期调控中的重要基因的影响,并对其机制进行初步探讨。方法:①运用四甲基偶氮唑盐(MTT)法等检测triptolide对CA46细胞株增殖的影响。②巢式甲基特异性PCR检测triptolide对CA46细胞p16基因甲基化模式的影响。③半定量RT-PCR检测经triptolide作用后CA46细胞p16基因mRNA的表达。结果:Triptolide对CA46细胞生长有明显的抑制作用,并有时间及剂量依赖性,48小时IC50为28.8ng/ml;triptolide可逆转P16基因的高甲基化;triptolide能够诱导CA46细胞p16基因mRNA的表达,具剂量依赖性。结论:小剂量triptolide可明显抑制CA46细胞的增殖,其可能机制为通过诱导CA46细胞中异常甲基化的p16基因去甲基化,使p16基因恢复表达,从而对细胞周期起负调控作用。

Objective：To observe the effects of Triptolide on proliferation of malignant lymphoma CA46 in vitro.Methods：The effects of Triptolide on proliferation CA46 cells were assayed by using cell culture,MTT.The effects of Triptolide on p16 gene of proliferation CA46 cells were analized by nested Methylation specific PCR and RT-PCR.Result：Following the treatment of triptolide,the cell proliferation rate was degraded as the treatment concentration increased and the culture time extended.The effects were dose and time-dependent.The 48 hour IC50 is 28.8ng/ml.Triptolide can reverse hypermethylation of p16 gene,and induce the expression of the mRNA.Conclusion：Small dose triptolide could depress the proliferation rate of CA46.The possible mechanism might be its reverseing the hypermethylation of P16 gene and activing the expression of P16 gene,to produce a con-regulation effect in cell cycle.