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目的:探讨桂枝茯苓丸诱导肿瘤细胞凋亡机制,为桂枝茯苓丸(GFW)开发应用提供实验依据。方法:计算抑瘤率,流式细胞仪测定细胞凋亡率,免疫组化法测定P21waf/cip蛋白表达,原位杂交法检测Survivin mRNA表达,电镜观察肿瘤细胞超微结构。结果:GFW抑瘤率为38.93%,流式细胞仪检测GFW组凋亡率17.79%,与模型组比较,差异有统计学意义(P<0.05)。GFW上调肿瘤细胞P21waf/cip蛋白表达,下调Survivin mRNA表达。GFW组镜下可见瘤细胞以凋亡变化为主。内膜结构完好,核膜清晰,细胞核固缩,染色质团块状散布核内或边集核膜下,并可见凋亡小体。结论:GFW诱导肿瘤细胞凋亡,其机制可能与上调P21及下调Survivin mRNA表达密切相关。
Objective: To investigate the mechanism of apoptosis of tumor cells induced by Guizhi Fuling Pills and provide experimental basis for the development and application of Guizhi Fuling Pill (GFW). Methods: The rate of tumor inhibition was calculated. Flow cytometry was used to determine the apoptosis rate. The expression of P21waf/cip protein was detected by immunohistochemistry. The expression of Survivin mRNA was detected by in situ hybridization. The ultrastructure of tumor cells was observed by electron microscope. Results: The inhibitory rate of GFW was 38.93%, and the apoptosis rate of GFW group was 17.79% by flow cytometry. Compared with the model group, the difference was statistically significant (P<0.05). GFW upregulated the expression of P21waf/cip protein in tumor cells and down-regulated the expression of Survivin mRNA. In the GFW group, tumor cells mainly showed apoptosis changes. The inner membrane structure is intact, the nuclear membrane is clear, the nucleus is pyknosis, the chromatin mass is scattered in the nucleus or the edge of the nuclear membrane, and apoptotic bodies are visible. CONCLUSION: The mechanism of GFW-induced apoptosis in tumor cells may be related to the up-regulation of P21 and down-regulation of Survivin mRNA expression.