消痰散结方对胃癌细胞系P16基因甲基化的影响

目的研究消痰散结方含药血清对胃癌细胞系基因甲基化的影响,探讨该方抗癌作用机制。方法培养胃癌细胞系MKN45、BGC823,制备消痰散结方药物血清。细胞分为空白对照组、空白血清组（无药大鼠血清）、药物血清组（消痰散结方药物血清）、5-Aza-CdR组（5-Aza-CdR）。CCK-8测定细胞活力,RT-PCR法测定P16基因表达,Methylight法测定基因P16甲基化水平。结果5-Aza-CdR和消痰散结方含药血清给药后72h后细胞增殖受到明显抑制,在MKN45细胞系中经过5-Aza-CdR和消痰散结方含药血清给药后72h后细胞增殖分别降低为76.14%和83.15%,在BGC823细胞系中经过5-Aza-CdR和消痰散结方含药血清给药后72h后细胞增殖分别降低为76.17%和83.44%,且消痰散结方药物血清组细胞增殖率明显低于5-Aza-CdR组。5-Aza-CdR和消痰散结方药物血清均能逆转P16基因甲基化,增加P16mRNA表达,与空白对照组比较有统计学差异（P<0.05）。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。

Effect of Xiaotan Sanjie Formula on DNA Methylation of P16 Gene in Gastric Cancer Cell Lines

Objective This study aimed to evaluate the effect of Xiaotan Sanjie Formula-contained serum on DNA methylation of P16 in gastric cancer cell lines to study its mechanism of anticancer.Methods Human gastric cancer cell lines,MKN45 and BGC823 were cultured and Xiaotan Sanjie Formula-containing serum was prepared.Cells were divided into four groups,i.e.blank control group,blank serum group,Xiaotan Sanjie Formula-containing serum group and 5-Aza-CdR group.Cell viability was determined by CCK8.The expression of P16 gene was detected by using RT-PCR.DNA methylation level of P16 gene was detected by Methylight method.Result Cell proliferation was obviously inhibited 72 hours after 5-Aza-CdR or Xiaotan Sanjie Formula-containing serum administered.In the MKN45 cell line,cell proliferation rate was decrease to 76.14% and 83.15% after treated with 5-Aza-CdR and Xiaotan Sanjie Formula-containig serum respectively;in the BGC823 cell line,the above indices were 76.17% and 83.44% respectively.The cell proliferation rate of Xiaotan Sanjie Formula-containing serum group was obviously lower than that of the 5-Aza-CdR group.Both drug containing serum could reverse methylation of P16 gene and increase expression of P16 mRNA.There was significant difference as compared with the blank control group （P<0.05）.Conclusion Xiaotan Sanjie Formula could suppress gastric cancer cell proliferation by reversing DNA methylation of P16 gene.