葫芦素B抑制视网膜母细胞瘤细胞生长的实验研究

Inhibitory effect of cucurbitacin B on the growth of retinoblastoma cells

目的研究葫芦素B对人视网膜母细胞瘤(RB)细胞凋亡的影响及作用机制。方法将体外培养的RB细胞分为对照组、低、中、高浓度葫芦素B组,MTT法测定细胞增殖率,Hoechst 33342/PI双染观察细胞凋亡,Annexin V/PI检测凋亡,镜观察细胞内结构改变,蛋白质印迹法检测Bax及Bcl-2蛋白相对表达量。结果葫芦素B剂量依赖性的抑制了RB细胞生长并诱导凋亡;葫芦素B干预后RB细胞胞浆浓缩,细胞器结构破坏,细胞核染色质凝结成块并固缩;蛋白质印迹结果显示,Bax蛋白表达增加,Bcl-2蛋白表达下降。结论葫芦素B通过改变线粒体凋亡途径中凋亡相关信号分子的表达,促进RB细胞凋亡。

Objective To study the effect and mechanism of cucurbitacin B on the apoptosis of retinoblastoma(RB)cells.Methods RB cells cultured in vitro were divided into four groups:the control group,and the low,medium and high concentration cucurbitacin B groups.Cell proliferation rate was measured by MTT method,cell apoptosis was observed by Hoechst 33342/PI double staining and cell apoptosis rate was detected by annexin V/PI.The changes in intracellular structure were observed by microscope,and the relative expression of Bax and Bcl-2 protein was detected by Western blot.Results Cucurbitacin B inhibited the growth of RB cells and induced apoptosis in a dose-dependent manner.After cucurbitacin B intervention,the cytoplasm of RB cells were concentrated,the organelle structure was damaged,and the nuclear chromatin was condensed.Western blotting showed that expression of Bax protein increased while the expression of Bcl-2 protein decreased.Conclusions Cucurbitacin B promotes RB cell apoptosis by changing the expression of apoptosis-related signal molecules in the mitochondrial apoptosis pathway.