消痰散结方对裸鼠胃癌原位移植瘤模型P16基因甲基化的影响

Effect of Xiaotansanjiefang on DNA Methylation of P16 in Nude Mouse Model of Human Gastric Carcinoma Cells Using Orthotopic Transplantation

目的观察消痰散结方对胃癌裸鼠原位移植瘤模型P16基因甲基化的影响,探讨消痰散结方抗癌作用机制。方法培养胃癌细胞系MKN-45,制备胃癌裸鼠原位移植瘤模型,分为模型对照组和中药组。中药组给予消痰散结方水煎剂(含原药材2.51g/mL),模型对照组给予生理盐水,连续6周。称重法测定抑瘤率,RT-PCR法测定P16基因表达,Methylight法测定P16基因甲基化水平。结果消痰散结方能抑制肿瘤生长,抑瘤率达54.82%。消痰散结方能逆转P16基因甲基化,增加P16mRNA表达,与模型对照组比较差异有统计学意义(P<0.01)。结论消痰散结方可以有效抑制胃癌细胞生长,逆转P16基因甲基化可能是其作用机制之一。

Objective To evaluate the effect of Xiaotansanjiefang on DNA methylation of P16 in the nude mouse model of human gastric carcinoma cells using orthotopic transplantation. Methods Twenty nude mouse models of human gastric carcinoma cell （MKN-45） were divided into two groups., control group and TCM group, treated with Xiaotansanjiefang （2.51 g/mL） and normal saline respectively. The tumor inhibition rate was measured by weighing method. The transcription level of P16 gene was detected by RT-PCR. DNA methylation level of P16 gene was detected by Methylight. Result The tumour weight was remarkably reduced after treated with Xiaotansanjiefang （54.82%）. The expression of P16 mRNA was remarkably increased and DNA methylation level of P16 gene was decreased in the gastric cancer cells of nude mice by Xiaotansanjiefang （P〈0.01）. Conclusion Xiaotansanjiefang can suppress cell proliferation by reversing DNA hypermethylation of P16 gene in human gastric cancer cell lines.