桑根白皮素调控人结肠癌HCT116细胞周期阻滞及其抑制细胞增殖体外研究.docVIP

桑根白皮素调控人结肠癌HCT116细胞周期阻滞及其抑制细胞增殖体外研究 　　摘要：目的 观察中药提取物桑根白皮素对结肠癌HCT116细胞增殖、细胞周期、细胞凋亡的影响，探讨其相关机制。方法 体外以不同浓度桑根白皮素作用于HCT116细胞72 h，CCK-8法检测细胞增殖，计算细胞生长抑制率和IC50，取25.4 μmol/L和50.8 μmol/L桑根白皮素作用于HCT116细胞24、48、72 h，CCK-8法检测细胞增殖，镜下观测细胞形态并摄图；25.4 μmol/L桑根白皮素作用于细胞24 h后，流式细胞仪检测细胞周期分布，荧光显微镜下TUNEL法原位检测细胞凋亡；Western blot检测药物干预后β-catenin通路蛋白和细胞周期蛋白的表达变化。结果 桑根白皮素对HCT116细胞增殖的抑制作用呈浓度/时间依赖性，72 h IC50为25.4 μmol/L；桑根白皮素阻滞HCT116细胞周期于G0/G1期和G2/M期，对细胞凋亡无明显诱导作用；桑根白皮素显著降低β-catenin通路相关蛋白β-catenin、c-Myc及细胞周期调控蛋白cyclinD1、cyclinB1的表达。结论 桑根白皮素可阻滞HCT116细胞周期，抑制细胞增殖，其机制可能是通过下调β-catenin通路活性，并抑制细胞周期蛋白cyclinD1、cyclinB1表达实现的。 　　关键词：桑根白皮素；结肠癌；细胞周期；细胞增殖；β-catenin通路 　　DOI：10.3969/j.issn.1005-5304.2015.05.020 　　中图分类号：R285.5 文献标识码：A 文章编号：1005-5304（2015）05-0072-04 　　Abstract：Objective To investigate the effects of Morusin on cell line proliferation， cell cycle and cell apoptosis of colorectal cancer HCT116 cell；To discuss its mechanism. Methods HCT116 cells were treated with different concentrations of Morusin for 72 h. Cell proliferation was detected by CCK-8 assay， and cell growth inhibition rate and IC50 value were calculated. HCT116 cells were treated with 25.4， 50.8 μmol/L Morusin for 24 h， 48 h and 72 h. Cell morphology was observed under the microscope， and cell proliferation was detected by CCK-8 assay. After HCT116 cell line was treated with 25.4 μmol/L Morusin for 24 h， cell cycle phase distribution was detected by flow cytometry and the cell apoptosis was detected by TUNEL assay under the fluorescence microscope. Post-intervention protein expressions were detected by Western Blot. Results The inhibitory effects of Morusin on the proliferation of HCT116 cell line was concentration/time dependent and IC50 value at 72 h was 25.4 μmol/L；Morusin induced the cell cycle arrest at G0/G1 phase and G2/M phase， but there was no induction of cell apoptosis；Morusin significantly decreased the expression of β-catenin and its target c-Myc， and downregulated the expressions of cyclinD1 and cyclinB1， which were invo