中药苦参碱对体外培养人宫颈癌Hela细胞抑制作用.docVIP

中药苦参碱对体外培养人宫颈癌Hela细胞抑制作用 　 作者：金畅，滕小玲，雷亚宁，周颖，郑膺，张婵 【摘要】 目的:探讨中药苦参碱对体外培养的人宫颈癌细胞株Hela细胞的抑制作用。方法:选用人宫颈癌Hela细胞进行体外培养，以0.125～2.0 mg/ml的苦参碱分别处理Hela细胞24～72 h后，MTT法检测细胞增殖，流式细胞仪（FCM）检测细胞凋亡率，透射电镜进行形态学观察，RT-PCR半定量检测凋亡相关基因Bcl-2和Bax的mRNA表达水平。结果:苦参碱对Hela细胞有抑制作用，呈剂量和时间依赖性, 其24 h半数抑制浓度（IC 50）为1.074 mg/ml；流式细胞仪检测到细胞在苦参碱作用后，细胞凋亡率增加；透射电镜观察发现部分细胞发生凋亡早期改变；RT-PCR结果显示凋亡相关基因Bcl-2/Bax比值随药物剂量增加而逐渐减少。结论:苦参碱对人宫颈癌Hela细胞具有明显抑制作用，诱导细胞凋亡可能是其机制之一。 【关键词】 苦参碱；宫颈癌；凋亡 Abstract: Objective:To explore the inhibitory effect of matrine on human cervical carcinoma Hela cells in vitro. Methods: The Hela cells were cultured in vitro and treated with different concentrations(0.125～2.0 mg/ml) of matrine solution for 24～72 hours. MTT assay was performed to evaluate the cytotoxin effect of matrine on Hela cells. FCM was used to detect the apoptosis of Hela cells. Transmission Electron Microscope analyses were carried out to examine the cell morphology. Semiquantitative RT-PCR was assessed to detect the mRNA levels of Bcl-2 and Bax gene expressions. Results: Different concentrations of matrines were cytotoxic to Hela cell in time-dependent and dose-dependent manner. The FCM analysis showed that the apoptosis rate increased after Hela cells were effected by matrine. Partial cells presented characteristic morphological changes under the transmission electron microscope. The results of RT-PCR showed that the rate of Bcl-2/Bax decreased when the matrine concentration increased. Conclusion: Matrine can inhibit the proliferation of human cervical carcinoma Hela cells in vitro. Inducing apoptosis may be one of the mechanisms. Key words: matrine; cervical carcinoma; apoptosis 苦参碱是从中药苦参（sophora flavescent ait）的干燥根中提取的具有广泛药理作用的活性物质，现代医学研究表明苦参碱具有抗病毒、抗感染、平喘、增强机体免疫力、升高白细胞、保肝等作用。最近研究表明,苦参碱在体外培养的肝癌、肺癌和白血病中有明显的抗肿瘤作用，且没有化疗药物的毒副作用[1-4]。因此，苦参碱的抗肿瘤作用引起广泛关注，成为研究的热点。但目前苦参碱对宫颈癌抑制作用方面的研究报道较少。本研究选择人宫颈癌Hela细胞，初步探讨苦参碱对体外培养的Hela细胞的抑制作用。 1 材料和方法 1.1 材料 宫颈癌细胞株Hela细胞由温州医学院第一附属医院科研中心惠赠。胎牛血清、DMEM培养基为Gibco公司产品。四甲基偶氮唑盐（MTT）、二甲基亚枫（DMSO）为美国Sigma公