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目的:观察不同浓度和作用时间二烯丙基二硫(DADS)对SW480细胞的生长抑制作用,应用前期蛋白质组学筛出的部分差异表达蛋白质分子,进行进一步分析与鉴定。方法:采用MTT、细胞计数法;施用流式细胞仪、RT-PCR和Western blotting方法,检测50μg/mL DADS对SW480细胞的凋亡率、CK19 mRNA和蛋白的表达水平。结果:MTT法显示,DADS能明显抑制SW480细胞生长增殖,呈浓度和时间依赖性,细胞生长增殖速度明显减慢。细胞计数表明,当DADS浓度增加时,SW480细胞群体倍增时间延长,与对照组比较,差异有统计学意义,P<0.05。流式细胞仪分析显示,50μg/mL DADS使SW480细胞凋亡率增加。RT-PCR和Western blotting结果证实,50μg/mL DADS处理SW480细胞后,CK19 mRNA和蛋白表达较对照组降低,差异有统计学意义,P<0.05。结论:DADS抑制SW480细胞系生长增殖,可能与CK19表达下调,促进细胞凋亡有关。
OBJECTIVE: To observe the inhibitory effect of diallyl disulfide (DADS) on the growth of SW480 cells at different concentrations and for different duration of action. Proteins differentially expressed in early proteomics were further analyzed and identified. Methods: The apoptotic rate and the expression of CK19 mRNA and protein of SW480 cells were detected by flow cytometry, RT-PCR and Western blotting using MTT assay and cell counting method. Results: MTT assay showed that DADS could significantly inhibit the growth and proliferation of SW480 cells in a concentration and time-dependent manner, and the proliferation and proliferation rate of SW480 cells was significantly slowed down. Cell counts showed that when DADS concentration was increased, doubling time of SW480 cell population was prolonged, which was significantly different from that of control group (P <0.05). Flow cytometry analysis showed that 50μg / mL DADS increased the apoptosis rate of SW480 cells. The results of RT-PCR and Western blotting confirmed that the expression of CK19 mRNA and protein in SW480 cells treated with 50μg / mL DADS was lower than that in control group, the difference was statistically significant (P <0.05). Conclusion: DADS can inhibit the growth and proliferation of SW480 cell line, which may be related to the down-regulation of CK19 and the promotion of cell apoptosis.