摘要
背景与目的:前期研究表明,二烯丙基二硫(diallyl disulfide,DADS)能有效在体内外抑制人结肠癌细胞增殖,并将细胞阻滞于G2/M期。本实验旨在进一步探讨DADS对人结肠癌SW480细胞周期阻滞作用的可能机制。方法:采用MTT、细胞计数法检测DADS对SW480细胞增殖的抑制作用;流式细胞术检测DADS对SW480细胞周期的影响;免疫细胞化学法检测DADS作用后PCNA、P53表达情况,蛋白印迹法分析P21WAF1、细胞周期蛋白B1(Cyclin B1)表达的改变。结果:不同浓度DADS作用SW480细胞24、48、72h后,对细胞增殖的抑制作用及细胞群体倍增时间呈浓度、时间依赖性增加。流式细胞仪分析结果显示,DADS作用SW480细胞后,G0/G1期细胞含量降低,S期细胞比例变化不大,而G2/M期细胞比例则明显增加,且随着处理浓度的增加和处理时间的延长,其G2/M期细胞含量逐渐增加,与对照组比较差异有统计学意义(P<0.05)。DADS作用后,免疫细胞化学检测显示,PCNA、P53蛋白表达明显下降。蛋白印迹分析显示,DADS作用SW480细胞后,Cyclin B1蛋白表达明显下降,P21WAF1蛋白的表达明显升高,均呈时间效应关系。结论:DADS可能通过下调PCNA、P53、Cyclin B1蛋白表达,上调P21WAF1蛋白表达引起SW480细胞G2/M期阻滞。
Background and Objective. Our previous study revealed that diallyl disulfide （DADS） significantly inhibited cell proliferation and induced cell cycle arrest at G2/M phase of human colon cancer SW480 cells. However, the molecular mechanism of ceil cycle arrest remains unclear. This study was to investigate the role and the molecular mechanism of DADS in the induction of cell cycle arrest of human colon cancer cell line SW480. Methods： Proliferation of SW480 cells after DADS treatment was measured by M-IF assay and cell counting. Phase distribution of cell cycle was analyzed by flow cytometry. Expression of PCNA, p53, p21WAF1 and cyclin B1 was detected by immunohistochemistry and Western blot. Results： DADS significantly inhibited proliferation and retarded the population doubling time of colonies in SW480 cells. DADS blocked cell cycle in the G2/M phase. Compared with the control group, SW480 cells treated with DADS markedly accumulated cells in the G2/M phase （P〈0.05）. Moreover, DADS remarkably decreased PCNA, p53 and cyclin B1 protein, but increased the expression of p21wF1 in a time and dose-dependent manner. Conclusion： DADS induces GJM arrest in human colon cancer SW480 cells, probably through the downregulation of PCNA, p53, cyclin B1 and upregulation of p21WAF1.
出处
《癌症》
SCIE
CAS
CSCD
北大核心
2009年第2期168-172,共5页
Chinese Journal of Cancer
基金
湖南省自然科学基金(No.02JJY2026)
湖南省教育厅课题(No.05C455
No.06C694)~~
