论文部分内容阅读
目的通过比较分析多西他赛(多西紫杉醇,docetaxel)对三阴性乳腺癌细胞系CAL-51和非三阴乳腺癌细胞系T47D的杀伤敏感性差异,探讨CAL-51对多西他赛耐药的可能机制。方法 MTT法测定不同剂量多西他赛对CAL-51和T47D细胞生长的影响并计算IC_(50)值;瑞氏-吉姆萨染色分析多西他赛作用后对两种细胞形态的变化;流式细胞仪(FCM)分析多西他赛对细胞周期的分布及凋亡情况的影响;荧光定量PCR检测分析多个基因在两种细胞中的差异表达;Western印迹法检测抗凋亡蛋白Bcl-2和胱天蛋白酶(caspase)家族蛋白在两种细胞中表达水平的差异。结果 T47D细胞经多西他赛处理后,形态出现显著变化;流式细胞检测显示,多西他赛能明显诱导非三阴乳腺癌细胞系T47D凋亡,与三阴性乳腺癌细胞系CAL-51相比具有显著性差异(P<0.01);定量PCR结果显示,CAL-51中抗凋亡基因Bcl-2高表达,与T47D相比具有显著性差异(P<0.05);蛋白质印迹法显示加药后两种细胞皆能活化内源性凋亡途径,但下游效应caspase的活化途径有所不同。结论多西他赛诱导2种细胞产生凋亡的内源性途径有所不同,Bcl-2的高表达可能是CAL-51细胞对多西他赛耐药的机制之一。
OBJECTIVE: To compare the sensitivity of docetaxel to triple-negative breast cancer cell line CAL-51 and non-triple-negative breast cancer cell line T47D and to compare the sensitivity of docetaxel against docetaxel Possible mechanism of medicine. Methods MTT assay was used to determine the effect of different doses of docetaxel on the growth of CAL-51 and T47D cells. IC 50 values were calculated. Wright-Giemsa staining was used to analyze the morphological changes of both cells after docetaxel treatment. FCM was used to analyze the effect of docetaxel on the cell cycle distribution and apoptosis. Fluorescent quantitative PCR was used to detect the differential expression of multiple genes in the two cell lines. Western blotting was used to detect the expression of Bcl- 2 and caspase family proteins in both cells. Results After treated with docetaxel, the morphology of T47D cells changed significantly. Flow cytometry showed that docetaxel could induce the apoptosis of non-triple-negative breast cancer cell line T47D obviously. Compared with triple-negative breast cancer cell line CAL-51 (P <0.01). The quantitative PCR results showed that the expression of anti-apoptotic gene Bcl-2 in CAL-51 was significantly higher than that in T47D (P <0.05) Both drugs can activate endogenous apoptotic pathway, but the downstream pathway of caspase activation is different. Conclusion Endogenous pathway of apoptosis induced by docetaxel is different. Bcl-2 overexpression may be one of the mechanisms of resistance to docetaxel in CAL-51 cells.