DADS抑制SW480细胞系生长增殖及CK19蛋白质分子鉴定

Proliferation inhibition effects by DADS in SW480 cell line and identification of CK19 protein

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 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.