马尾松树皮提取物体外抑制人大肠癌细胞生长机理初探

为探讨马尾松树皮提取物(PMBE)体外抑制大肠癌LoVo细胞生长的作用特点和机理,通过MTT、显微镜术、凝胶电泳和流式细胞术,研究PMBE抑制LoVo细胞生长的特点,运用免疫组化和RT-PCR探究相关分子机理。结果表明:PMBE时间-剂量依赖地抑制LoVo细胞的体外生长;PMBE处理后,流式细胞检测发现LoVo细胞周期阻滞于G1或S期,同时可检测到亚二倍体峰的出现;荧光镜检和透射电镜观察可看到LoVo细胞的核皱缩、边集甚或碎裂,以及有凋亡小体形成,其基因组经过凝胶电泳呈现典型的梯形Ladder;RT-PCR和免疫组化结果显示PMBE处理可上调LoVo细胞中p53和p21基因的转录效率,并下调Bcl-2的蛋白表达量。说明PMBE通过上调p53和p21的表达量阻滞细胞周期、下调Bcl-2的表达量诱导细胞凋亡的双重机制,抑制LoVo细胞的体外生长,可供进一步探索相关信号传导通路参考。

Primary exploration of mechanism of Pinus massoniana bark extract inhibiting growth of human colorectal carcinoma cells in vitro

To explore the character and mechanism of Pinus massoniana bark extract(PMBE) inhibiting the growth of human colorectal carcinoma LoVo cells in vitro.MTT,microscopy,gel electrophoresis and flow cytometry were used to identify the characters of PMBE inhibiting LoVo cell growth and inducing apoptosis.Immunohistochemistry analysis and RT-PCR were used to explore the related molecular mechanism.PMBE inhibits the growth of LoVo cells in a time-and dose-dependent manner.PMBE treatment resulted in cell cycle arrest at G1/S phase,Sub-G1 curve,genomic DNA ladder pattern,and morphological changes,including nuclear condensation,boundary aggregation or split,and apoptotic body formation.In addition,PMBE up-regulates the expression of p53 and p21 and down-regulates the expression of Bcl-2.PMBE inhibits the growth of LoVo cells in vitro via double mechanisms of cell cycle arrest through up-regulation of p53 and p21 expression and apoptosis induction through down-regulation of Bcl-2 expression,which laid foundation for further research of the related signal transduction pathways.