二烯丙基二硫对人鼻咽癌CNE2细胞周期的阻滞作用

Arrest effect of diallyl disulfide on cell cycle in human nasopharyngeal cancer CNE2 cells

目的探讨二烯丙基二硫(DADS)对人鼻咽癌CNE2细胞周期的阻滞作用及分子机制。方法体外培养CNE2细胞,采用MTT比色实验、细胞计数法、细胞形态学观察、流式细胞仪和Western blotting方法分析DADS对CNE2细胞的增殖抑制作用、细胞周期分布的影响及细胞周期相关蛋白p21WAF1的表达。结果MTT法显示,不同浓度DADS(90、140、240、400μmol/L)处理CNE2细胞48小时后,生长抑制率分别为3.3%、12.9%、28.3%、56.9%;细胞计数法表明,常规培养CNE2细胞群体倍增时间为24.5小时,DADS的浓度由90μmol/L增加到400μmol/L时,其细胞群体倍增时间由27.6小时延长到93.1小时;HE染色结果显示,不同浓度DADS处理CNE2细胞48小时后细胞异型性明显减少,核浆比例明显减少;流式细胞仪分析显示DADS呈浓度依赖性将CNE2细胞阻滞在G0/G1期;Western blotting分析表明,在细胞周期阻滞的同时有p21WAF1蛋白表达上调。结论DADS对CNE2细胞的抑制增殖作用与G0/G1期阻滞有关,其分子机制可能与调节p21WAF1表达有关。

Objective To investigate the arrest effect of diallyl disulfide （DADS） in the cell cycle of human nasopharyngeal carcinoma CNE2 cell line and its molecular mechanism. Methods The growth inhibition of CNE2 cell line was measured by MTT assay and cell counting. Phase distribution of cell cycle was analyzed by flow cytometry. The expression of p21WAF1 was determined by Western blotting analysis. Results MTT assay showed that DADS inhibited growth of CNE2 cells significantly and exhibited a dose dependent modal. Adding 90 μmol/L, 140μmol/L, 240 μmol/L and 400μmol/L DADS for 48 hours suppressed CNE2 growth by 3. 3%, 12. 9%, 28. 3% and 56. 9%, respectively. Cell counting showed that average doubling time retarded from 24. 5 hours in normal cultured CNE2 cells to 93. 1 hours in 400 μmol/L DADS experimented CNE2 cells （P〈 0. 05）. Flow cytometry analysis revealed that CNE2 cells treated with increasing quantities of DADS increased the percentage of cells in the G0/G1 phase. Western blotting analysis suggested that p21WAF1 was up-regulated when treated with DADS in a dose dependent manner. Conclusion The antiproliferation property of diallyl disulfide （DADS） in cultured human nasopharyrtgeal carcinoma CNE2 cell line relates to its ability to arrest G0/G1 through over-expression of p21WAF1.