桂枝茯苓胶囊对体外人宫颈癌Hela细胞抑制作用及机理研究(1)
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收稿日期:2009-11-06
基金项目:浙江省教育厅基金资助项目(20060721)
作者简介:黄燕芬(1974-),女,浙江金华人,副教授,主要从事中药成分的分离与中药药理研究。
摘 要:目的:研究桂枝茯苓胶囊对体外培养人宫颈癌Hela细胞的增殖抑制作用和作用机理的初步研究。方法:MTT比色法观察桂枝茯苓胶囊对人宫颈癌Hela细胞增殖的抑制作用;免疫组化检测Fas、Fas –L、Bcl-2基因的表达;western-blot和RT-PCR检测对人宫颈癌Hela细胞Caspase-3蛋白和Caspase-3 MRNA的表达影响;DNA-Ladder检测凋亡条带。结果:桂枝茯苓胶囊对人宫颈癌Hela细胞有明显的抑制作用(P<0.01);桂枝茯苓胶囊能促进Fas、Fas –L表达,抑制Bcl-2的表达;能增加Hela细胞Capase-3的蛋白表达;琼脂糖凝胶电泳中呈现典型的“阶梯状”图谱。结论:桂枝茯苓胶囊有体外抗肿瘤作用,其机制可能是通过促进Fas、Fas –L表达、抑制bcl-2的表达;增加Capase-3的表达实现的。
关键词:桂枝茯苓胶囊;宫颈癌Hela细胞;MTT;western-blot;免疫组化
中图分类号:R285.5 文献标识码:A 文章编号:1673-7717(2010)04-0774-04
Study of Inhibition and Mechanism on Hela of Guizhi Fuling Capsule in Vitro
HUANG Yanfen,DONG Gaixia,CHAI Hui, YUAN Xiaofeng
(Life Science Institute,Zhejiang Chinese Medical University, Hangzhou 310053, Zhejiang,China)
Abstract:Objective: To study the effects on anti-proliferation and its mechanism on the Hela of Guizhifuling Capsule. Methods: The inhibitory effect on Hela cell proliferation was assayed by MTT. Fas,FasL,Bcl-2 gene's expression was detected by Immunohistochemistry. caspase-3 MRNA and protein were detected by Western-blot and RT-PCR. Results: Guizhifuling Capsule have obvious effect on anti-proliferation of Hela cells (P<0.01). Guizhifuling Capsule can promote Fas and Fas –L to express and inhibit expression of Bcl-2. Guizhifuling Capsules can reduce expression of capase-3 protein and promote capase-3 MRNA in Hela. Typ icalDNA ladder appeared in agarose gel electrophoresis.Conclusion: Guizhifuling Capsule can inhibit the inhibit proliferation of Hela cells. Promoting the expression of caspase-3, promoting the expression of Fas expression and inhibiting the expression of Bcl-2 may be the mechanisms of its anti-tumor.
Key words:Guizhifuling capsule; Hela; western-blot; immunohistochemistry; RT-PCR
宫颈癌是发生在全球妇女中仅次于乳腺癌的第2个常见的恶性肿瘤[1]。临床上治疗常用化疗药物,化疗药物副作用大,容易产生耐药性。中药治疗肿瘤已经越来越受医务人员和患者青睐。
桂枝茯苓胶囊主要由桂枝、茯苓、桃仁、丹皮、赤芍等5味药物组成。具有活血化瘀、消症散、抗凝血、促进微循环、抗肿瘤等多种功效,现广泛用于保守治疗子宫肌瘤[2-3]。
本实验旨在观察桂枝茯苓胶囊对体外培养人宫颈癌Hela细胞生长抑制作用,从诱导凋亡调控基因角度分析其抑制作用机制,为深度开发利用桂枝茯苓胶囊提供实验依据 ......
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