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目的:探讨5-氮杂-2’-脱氧胞苷(5-aza-2’-deoxycytidine,5-Aza-dC)对肿瘤多药耐药细胞生长抑制率、细胞周期的影响及可能的作用机制。方法:采用MTT法检测5-Aza-dC对人乳腺癌耐药细胞株MCF-7/ADR、口腔表皮癌多药耐药细胞KBV/200及其相应的亲本敏感MCF-7和KB细胞的生长抑制率;FCM法检测不同浓度5-Aza-dC(1、5和10μmol/L)作用72h及5-Aza-dC(10μmol/L)作用24、48和72h后,各细胞周期分布的变化;分别采用实时荧光定量-PCR及蛋白质印迹法检测细胞周期相关蛋白cyclin A、cyclin E和p21waf1/cip1mRNA及蛋白的表达情况。结果:5-Aza-dC对耐药细胞株MCF-7/ADR及KBV/200均呈现明显的生长抑制作用,细胞周期被阻滞在G2/M期,并呈时间及剂量依赖性;细胞周期相关蛋白cyclin A、cyclin E和p21waf1/cip1mRNA及蛋白的表达均上调。结论:5-Aza-dC可以通过使MCF-7/ADR及KBV/200细胞的细胞周期被阻滞在G2/M期,从而抑制肿瘤多药耐药细胞的增殖;其作用机制可能与上调细胞周期相关蛋白cyclin A、cyclin E和p21waf1/cip1的表达有关。
Aims: To investigate the effect of 5-aza-2’-deoxycytidine (5-Aza-dC) on the growth inhibition and cell cycle of multidrug-resistant tumor cells and its possible mechanism . Methods: MTT assay was used to detect the growth of multi-drug resistant cell line MCF-7 / ADR, oral multi-drug resistant cell line KBV / 200 and its corresponding parental MCF-7 and KB cells in vitro (FCM) was used to detect the changes of cell cycle distribution at different concentrations of 5-Aza-dC (1, 5 and 10μmol / L) for 72h and 5-Aza-dC The expressions of cyclin A, cyclin E and p21waf1 / cip1 mRNA and protein were detected by real-time fluorescence quantitative PCR and Western blot respectively. Results: 5-Aza-dC showed obvious growth inhibitory effects on MCF-7 / ADR and KBV / 200 cell lines, and the cell cycle was arrested in G2 / M phase in a time and dose dependent manner. Cell cycle The expressions of cyclin A, cyclin E and p21waf1 / cip1 mRNA and protein were up-regulated. CONCLUSION: 5-Aza-dC can inhibit the proliferation of multidrug-resistant tumor cells by blocking the cell cycle of MCF-7 / ADR and KBV / 200 cells in G2 / M phase. The mechanism may be related to the upregulation of cells Cyclin A, cyclin E and p21waf1 / cip1 were correlated with the expression of cyclin A and cyclin E.