胃热消炎合剂对人胃癌细胞SGC-7901增殖影响的机制研究

Study on the effect of Weire Xiaoyan mixture on proliferation of human gastric cancer cell SGC-7901 and its mechanism

目的研究胃热消炎合剂对人胃癌细胞SGC-7901增殖的影响。方法取对数生长期的人胃癌细胞SGC-7901进行体外实验,其中空白血清组加入含10%正常血清培养,低剂量含药血清组加入含2.5%药物血清+7.5%正常血清培养,中剂量含药血清组加入含5%药物血清+5%正常血清培养,高剂量含药血清组加入含10%药物血清培养,胃热消炎合剂含药血清通过给大鼠灌胃的方式获得。分别采用CCK-8法、EdU染色法和免疫荧光法检测各组细胞增殖情况,采用RT-PCR法检测各组细胞增殖信号通路p53-p21的相关基因p21、p53、CyclinD1、CDX2、CDK7mRNA的表达情况,采用Western blot法检测各组细胞中p21、p53、CyclinD1、CDX2蛋白表达水平。结果各含药血清组SGC-7901细胞增殖受到明显抑制,且呈浓度依赖性。各含药血清组p21、p53 mRNA表达水平均明显高于空白血清组(P均<0.05),CyclinD1 mRNA表达水平均明显低于空白血清组(P均<0.05),且呈浓度依赖性;各组CDX2、CDK7mRNA表达水平比较差异均无统计学意义(P均>0.05)。各含药血清组p21蛋白表达水平均明显高于空白血清组(P均<0.05),且呈浓度依赖性;低剂量含药血清组和中剂量含药血清组p53蛋白表达水平均明显高于空白血清组(P均<0.05),而高剂量含药血清组p53蛋白表达水平明显低于空白血清组(P<0.05);各组CyclinD1、CDX2蛋白表达水平比较差异均无统计学意义(P均>0.05)。结论胃热消炎合剂可能通过作用于细胞周期来抑制人胃癌细胞SGC-7901增殖,其可能具有预防和治疗早期胃癌的作用。

Objective It is to study the effect of Weire Xiaoyan mixture on the proliferation of human gastric cancer cell line SGC-7901.Methods Human gastric cancer cells SGC-7901 in logarithmic growth phase were used for the experiment in vitro.The blank serum group was cultured with 10%normal serum,the low-dose drug-containing serum group was added with 2.5%drug serum+7.5%normal serum,and the medium-dose drug-containing serum group was added with 5%drug serum+5%normal serum,the high dose-containing serum group was cultured by adding 10%drug-containing serum,and the Weire Xiaoyan mixture drug-containing serum was obtained by intragastric administration to rats.CCK-8 method,EdU staining method and immunofluorescence method were used to detect the cell proliferation in each group,and RT-PCR method was used to detect the expression of correlated genes of p53-p21 proliferation signal pathway p21,p53,CyclinD1,CDX2 and CDK7 mRNA,Western blot was used to detect the expression levels of p21,p53,CyclinD1 and CDX2 protein in the cells of each group.Results The proliferation of SGC-7901 cells in each drug-containing serum group was significantly inhibited,and it was concentration-dependent.The levels of p21 and p53 mRNA expression in each drug-containing serum group were significantly higher than those in the blank serum group(all P<0.05),and the levels of CyclinD1 mRNA expression were significantly lower than those in the blank serum group(all P<0.05),and they were concentration-dependent;There were no significant differences in the expression levels of CDX2 and CDK7 mRNA between every groups(all P>0.05).The levels of p21 protein expression in each drug-containing serum group were significantly higher than those in the blank serum group(both P<0.05)and they were concentration-dependent;the levels of p53 protein expression in the low-dose drug-containing serum group and the medium-dose drug-containing serum group were significantly higher than that in the blank serum group(all P<0.05),but the p53 protein expression in high-dose drug-containing serum group was significantly lower than the blank serum group(P<0.05);there was no significant difference in the expression levels of CyclinD1 and CDX2 protein between each group(P>0.05).Conclusion Weire Xiaoyan mixture may inhibit the proliferation of human gastric cancer cell SGC-7901 by acting on the cell cycle,which may have the effect of preventing and treating on early gastric cancer.