桂枝茯苓丸对荷瘤鼠P21^(waf/cip)基因表达的影响

Guizhi Fuling Wan's effect on the expression of P21^(waf/cip) in bearing tumor mice

目的探讨桂枝茯苓丸诱导肿瘤细胞凋亡的机制,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P21waf/cip及下调Survivin mRNA表达密切相关。

Objective To study the effect of Guizhi Fuling Wan （GFW） on tumor apoptosis and apply the experimenting basis of GFW＇s development and utilization. Methods The S180 mice sarcoma model was used to detect the inhibitory effects of GFW, observe the ultrastructural change by electron microscopy, and the flow cytometer was used for apoptosis determination. The protein expression of P21^waf/cip and the mRNA expression of Survivin were evaluated by immunohistochemistry and in situ hybridization. Results For S180 sarcoma, the inhibition ratio of GFW was 38.93%. The apoptosis was 17.79% by the flow cytometer. Some typical apoptotic cells and apoptotic bodies were observed through electron microscope. Chromatin gathered at the side of cell, the nucleus turned into the nucleus band and nucleus projected. Survivin mRNA markedly declined in the GFW group when compared with that in the model group （P〈0.01）, while the postitive expression of P21^waf/cip was obviously higher in the GWF group than in the model group （P〈0.01）.Conclusion GFW induces tumor cell apoptosis, the mechanism of which may be closely related to adjusting expression level of P21^waf/cip and Survivin gene.