黄芪注射液对宫颈永生化上皮细胞生长的作用

Effect of Huangqi Injections on Growth of Human Immortalized Cervical Epithelial Cells

目的:观察黄芪注射液对人宫颈永生化上皮细胞(H8细胞)的生长抑制作用,分析其细胞周期调控机制。方法:采用MTT法检测黄芪注射液对H8细胞生长的作用,光学显微镜观察药物作用后细胞形态,利用流式细胞仪(FCM)分析黄芪注射液作用后H8细胞的周期。免疫荧光分析的第一抗体为细胞周期蛋白(cyclin)Dl,B1,细胞周期蛋白依赖性激酶(CDK)1,4和细胞周期蛋白依赖性激酶抑制因子(CDKI)p21蛋白,第二抗体采用荧光素标记,应用FCM检测黄芪注射液作用后H8细胞荧光强度。结果:黄芪注射液对H8细胞的生长有抑制作用,其中20 g·L-1作用最明显。黄芪注射液处理组可见细胞生长稀疏、死亡或凋亡,细胞固缩成深染圆形小体。黄芪注射液作用于H8细胞时,S,G2期细胞减少,使细胞阻滞于G1期。H8细胞cyelin D1蛋白表达降低极显著;cyclin B1,CDK4蛋白,p21蛋白表达略降低,无统计学意义;CDK1蛋白表达显著降低。结论:黄芪注射液有可能作为一种细胞周期调控剂来预防或辅助治疗宫颈癌前病变。

Objective： To observe growth inhibition of Huangqi injections on human immortalized cervical epithelial cells （H8 cells） and make research on cell cycle regulation mechanism. Method： Effect of Huangqi injections on H8 cells was detected by MTT, form of cells after being injected was observed on optical microscope, cells cycle of H8 was analyzed by flow cytometry （FCM）. First antibodies of immunofluorescenee assay were cyclins D1 and B1, cyclin-dependent kinases （CDK） 1 and 4, cyclin-dependent kinase inhibitor （p21）, second antibody was fluorescein labeled, fluorescence degree of H8 cells after being injected Huangqi injections was determined by FCM. Result： There was inhibiting effect on growth of H8 cells after being injected, function was the most obvious of 20 g·L^-1 Some of the Huangqi injections treatment group appeared to he sparse in cell growth, death or apoptosis, cells stained pyknosis hyperchromatic circular corpuscle. When Huangqi injections on H8 cells, S and G2 phase cells decreased, cell blocked in G1 period. Expression of cyelin D1 protein in H8 cells significantly decreased; expression of cyclin B1, CDK4 and P21 were slightly reduced, but without statistical significance; expression of CDK1 protein reduced significantly. Conclusion： Huangqi injections can be regarded as a cell cycle regulation agent for prevention or adjuvant therapy of cervical cancer.