【作者】 崔映宇； 王宏斌； 谢衡； 伍春莲； 彭青青； 郑光耀； 王金发；

【Author】 CUI Ying-yu 1, WANG Hong-bin1, XIE Heng 2, WU Chun-lian 1, PENG Qing-qing 1, ZHENG Guang-yao2, WANG Jin-fa 1* (1. State Key Laboratory for Biocontrol and The Key Laboratory of Gene Engineering of Ministry of Education, College of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China; 2. Institute of Songzhen Nutritional Resource, Guangzhou 510645, China)

【机构】 生物防治国家重点实验室教育部基因工程重点实验室中山大学生命科学院； 广州嵩珍营养源研究所；

【摘要】 目的探讨PMBE体外抑制LoVo细胞生长的作用特点和机理。方法通过MTT、显微镜术、凝胶电泳和流式细胞术研究PMBE抑制细胞生长的特点和凋亡鉴定,运用免疫组化和RT-PCR探究相关分子机理。结果PMBE能时间-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,下调Bcl-2的蛋白表达。结论PMBE通过上调p53和p21阻滞细胞周期和下调Bcl-2诱导细胞凋亡双重机制抑制LoVo细胞的体外生长,为进一步探索相关信号传导通路奠定了基础。更多还原

【Abstract】 Aim To explore the character and mechanism of Pinus massoniana bark extract (PMBE) inhibiting the growth of human colorectal carcinoma LoVo cells in vitro. Methods 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. Results PMBE inhibit 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. Conclusion 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.更多还原