【摘要】 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。更多还原

【Abstract】 OBJECTIVE:To investigate the arrest effect of diallyl disulfide (DADS) in the cell cycle of human nasopharyngeal carcinoma SW480 cell line and its molecular mechanism. METHODS: The growth inhibition effect of different concentrations of DADS (50 μg/mL) on SW480 cell line was measured by MTT assay and cell counting. Apoptosis rate was analyzed by flow cytometry. CK19 mRNA level was detected by RT-PCR. The expression of CK19 was determined by Western blotting analysis. RESULTS: The MTT assay showed that DADS inhibited growth of SW480 cells significantly with dose dependent. There was a significant difference between the treated and controlled cases (P<0.05). Cell counting results showed that average doubling time increased with the dose increase of DADS, compared with normal cultured SW480 cells (P<0.05). Flow cytometry analysis revealed that SW480 cells treated with 50 μg/mL DADS increased the apoptosis rate. CK19 mRNA was decreased in SW480 treated with 50 μg/mL DADS using RT-PCR. CK19 was decreased significantly in SW480 treated with 50 μg/mL DADS by using Western blotting. The differences were more than two timesc. CONCLUSION: The anti-proliferation property of DADS in cultured human colon carcinoma SW480 cell line may relate to its ability of increasing the apoptosis rate through down-regulation of CK19.更多还原