苦参碱抑制视网膜母细胞瘤细胞增殖的研究

Study on the mechanism of inhibition of matrine on the proliferation of retinoblastoma cells

目的探讨中药成分苦参碱(Ma)对视网膜母细胞瘤细胞株(HXO-Rb44细胞株)增殖的抑制作用及相关作用机制,为中药综合治疗视网膜母细胞瘤提供实验依据。设计实验性研究。研究对象体外培养的HXO-Rb44细胞。方法0.4mmol/L的Ma分别作用HXO-Rb44细胞12、24、48小时,流式细胞仪检测S期细胞百分比的变化;通过细胞免疫组织化学染色结合计算机自动图像分析技术,测量阳性细胞积分光密度(IOD)值,观察分析Ma对HXO-Rb44细胞内增生靶基因PCNA的调控。主要指标S期细胞百分比(SPF)、积分光密度值(IOD)。结果0.4mmol/L浓度Ma作用HXO-Rb44细胞后,S期细胞百分比下降(P均<0.01),细胞内PCNA蛋白表达水平下降(P均<0.01),并均呈时间依赖性。结论Ma能够抑制HXO-Rb44细胞增殖,提示Ma有望成为治疗视网膜母细胞瘤的有效药物。

Objective To evaluate the effects of matrine on retinoblastoma cells in vitro, and to explore the possible molecule mechanisms of its inhibition on the proliferation of retinoblastoma cell. Design Experimental study. Participants Cultured HXO-Rb44 cells in vitro. Methods After the cells were treated with matrine for various durations, S-peried frequency（SPF） was evaluated with flow cytometry. The expressions of proliferating cell nuclear antigen （PCNA） were analyzed with immunohistochemical stain technique. The positive results were assessed with integral optical density （IOD） in computer image analysis system. Main Outcome Measures SPF, IOD. Results Matrine down-regulated the SPF and inhibited the expression of PCNA in HXO-Rb44 cells at protein levels. The inhibition showed a time-dependent effect. Conclusion Matrine can inhibit the proliferation of retinoblastoma cells in a time-dependent manner. The effect associates with its down-regulating the expression of PCNA in retinoblastoma cells, which may be one of important mechanisms. It suggests that matrine may be a potent drug for the treatment of retinoblastoma.