文档介绍：Calphostin C对乳腺癌细胞增殖的抑制作用及相关机制
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作者:千新来,崔静,任峰,赵杰,冶亚平,贺国洋,李新强,朱慧芳,韩芳毅,杨慧林,王霞,原志庆
【摘要】目的研究Calphostin C对乳腺癌细胞MDAMB435S增殖的影响及初步机制。方法通过观察Calphostin C处理MDAMB435S细胞前后细胞的生长速度、倍增时间、克隆形成率和细胞周期分布等变化,探讨Calphostin C对MDAMB435S细胞增殖的影响;通过Western blotting、免疫细胞化学染色和RTPCR等方法探讨Calphostin C影响MDAMB435S细胞增殖的初步机制。结果与MDAMB435S亲本细胞(对照组)相比,Calphostin C处理的MDAMB435S细胞(实验组)生长速度明显减慢,倍增时间明显延长(),并且呈现浓度和时间依赖性;对照组和实验组细胞克隆形成率分别为(±)%和(±)%,差异有统计学意义();实验组细胞周期出现明显的G1期阻滞()。Western blotting和免疫细胞化学染色结果显示,与对照组细胞相比,实验组细胞中蛋白激酶C
α(protein kinase Cα, PKCα)表达没有明显变化,但活性状态(胞质转位至胞膜)的PKCα显著减少。Western blotting和RTPCR结果显示,与对照组细胞相比,实验组细胞中p53、Cyclin A和Cyclin B表达没有明显变化,但p21表达上调,Cyclin E表达下调。结论 Calphostin C可显著抑制乳腺癌细胞MDAMB435S的增殖,该作用可能与其抑制PKCα活性、上调p21表达和下调Cyclin E表达有关。
【关键词】钙磷酸蛋白C;蛋白激酶C;乳腺癌;增殖
China)ABSTRACT: Objective To explore the effects of Calphostin C on the proliferation of breast carcinoma cells (MDAMB435S) and its preliminary mechanism. Methods The effects of Calphostin C on the proliferation of MDAMB435S cells were studied by observing changes of the growth rate, doubling time, cloneforming efficiency, and distribution of cell cycle of MDAMB435S cells before and after Calphostin C treatment. Furthermore, the mechanism involved was analyzed by Western blotting, immunocytochemistry and RTPCR assay. pared with those of the parental MDAMB435S cells (control group), the growth rate was suppressed markedly and the doubling time was prolonged significantly () in the MDAMB435S cells treated with Calphostin C (experimental group), and the changes presented concentration
 and timedependence patterns. Furthermore, the cloneforming efficiency in control group and experimental group was (±)% and (±)%, respectively, with a significant difference (). Moreover, for the experimental group cells, the cell cycle arrested in G1 phase (). The results from Western blotting and immunocytoch