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目的:观察健脾解毒散结中药胃康宁含药血清对胃癌细胞周期调控因子CyclinD1,CyclinD2,Cdk4(Cyclin dependant kinase4,CDK4),Cdk6和p16INK4amRNA及其蛋白表达的作用。方法:分别将高、中、低不同剂量的中药胃康宁制剂igSD大鼠,制备含药血清,然后分别用不同剂量含药血清培养胃癌细胞;采用免疫组织化学SABC法及RT-PCR法检测各组胃癌细胞中CyclinD1,CyclinD2,Cdk4,Cdk6和p16INK4amRNA和蛋白的表达。结果:免疫组化结果显示,与空白对照组比较,胃康宁高、中、低剂量组CyclinD1灰度值变化,无统计学意义(P>0.05);CyclinD2,Cdk4,Cdk6灰度值均有明显升高,而p16INK4a灰度值则显著降低(P<0.05,P<0.01);RT-PCR结果显示,胃康宁高、中、低剂量组CyclinD1 OPTDI值变化,无统计学意义(P>0.05);但能明显降低CyclinD2,Cdk4和Cdk6 OPTDI值,同时升高p16INK4a OPTDI值(P<0.05,P<0.01)。结论:中药胃康宁可抑制胃癌细胞促细胞周期相关因子CyclinD2,Cdk4和Cdk6的表达,同时增强胃癌细胞周期抑制因子p16INK4a的表达,这可能是健脾解毒散结中药胃康宁抑制胃癌细胞生长的作用机制。
Objective: To observe the effect of medicated serum containing Weipi Sanjie Sanjie Decoction on cell cycle regulatory factors CyclinD1, CyclinD2, Cdk4 (CyclinD3K4), Cdk6 and p16INK4amRNA and their protein expression in gastric cancer. Methods: Weigang, middle and low doses of traditional Chinese medicine Weikang Ning preparation igSD rats, preparation of drug-containing serum, and then with different doses of drug-containing serum cultured gastric cancer cells; using immunohistochemical SABC method and RT-PCR assay The expression of CyclinD1, CyclinD2, Cdk4, Cdk6 and p16INK4amRNA and protein in gastric cancer cells. Results: The results of immunohistochemistry showed that compared with the blank control group, the gray value of CyclinD1 in Weichengling group had no significant difference (P> 0.05), and the gray value of CyclinD2, Cdk4, (P <0.05, P <0.01). The results of RT-PCR showed that there was no significant difference in the OPTDI value of CyclinD1 between the high, medium and low doses of Weichengning (P> 0.05) ; However, the OPTDI values of CyclinD2, Cdk4 and Cdk6 were significantly decreased and the OPTDI value of p16INK4a was also increased (P <0.05, P <0.01). Conclusion: Weitangning can inhibit the expression of cell cycle-related factors CyclinD2, Cdk4 and Cdk6 and enhance the expression of cell cycle inhibitor p16INK4a in gastric cancer cells, which may be the effect of traditional Chinese medicine Weipeikang, which inhibits the growth of gastric cancer cells mechanism.