摘要
目的探讨芪三酚(Res)对人乳腺癌MDA-MB-231细胞增殖抑制的相关效应及其与MDC1基因的关系。方法以人乳腺癌MDA-MB-231细胞株为研究对象,采用MTS方法测定细胞增殖,应用吖啶橙荧光染色观察Res对乳腺癌MDA-MB-231细胞的影响,用RT-PCR与免疫印迹方法测定MDC1基因与蛋白表达水平,用小RNA干扰MDC1基因后,用流式细胞仪检测细胞的凋亡并观察其对Res的敏感性影响。结果 40μmol/L以上的Res可显著抑制乳腺癌MDA-MB-231细胞的增殖(P<0.05),给予0、60、120μmol/L Res能明显降低MDC1基因和蛋白的表达(P<0.05)。用小RNA干扰MDC1基因后,流式细胞术分析显示,实验组(MDC1-siRNA)的细胞凋亡率[(45.13±6.2)%]较阴性对照组[(24.34±2.6)%]和未处理组[(17.69±4.9)%]明显上升(P<0.05),MTS结果显示MDC1基因干扰后细胞对Res的敏感性增加。结论40μmol/L以上的Res可以抑制MDA-MB-231细胞的增殖,Res可以有效降低MDC基因和蛋白的表达并促进细胞的凋亡。用小RNA干扰MDC1基因(MDC1-siRNA)后,MDA-MB-231细胞对Res的敏感性增加。
Objective To investigate the effect of resveratrol（Res） on the proliferation of human breast cancer cell line MDA- MB-231 and expression of MDC1 gene. Methods Breast cancer cell MDA-MB-231 was used for the study. The proliferation of breast cancer cell MDA-MB-231 was measured by MTS assay. The MDA-MB-231 cell apoptosis was observed by acridine orange fluorescent staining. The expressions of MI）C1 gene and protein were tested by PCR （qRT-PCR）and Western blot, respectively. The MDA-MB-231 cell apoptosis was determined by flow cytometry. The effect of resveratrol and small interfering RNA interference （siRNA） MDC 1 gene knockdown on cell proliferation and apoptosis were also measured by flow cytometry. Results MTS assay showed the proliferation of breast cancer cell MDA-MB-231 could be significantly inhibited by 50 p^mol/L and above of Res （P〈O.05）. Acridine orange fluorescent staining showed Res enhanced the apoptosis and autophagy of MDA- MB-231 cells. PCR（qRT-PCR）and Western blot showed the expression of MDC1 gene and protein could be decreased by 80 and 160 pLmol/L Res （P〈0.05）. Flow cytometry showed the apoptotic rate of experimental group with MDCI-siRNA knockdown was （45.13＋6.2）%, while the apoptotic rate of negative control group was （24.34_＋2.6）% and that of without treatment group was （17.69~4.9）%. MDC1 knockdown by siRNA, could also enhance the drug sensitivity of MDA-MB-231 cells to resveratrol. Conclusion Resveratrol could inhibit the proliferation of MDA-MB-231 cells and decrease the expression of MDC1 gene and protein. MDCI-siRNA gene knockdown could significantly decrease the apoptosis of MDA-MB-231 cells and increase the sensitivity of MDA-MB-231 ceils to resveratrol.
出处
《热带医学杂志》
CAS
2013年第11期1319-1323,F0003,共6页
Journal of Tropical Medicine
基金
高校博士学科点专项科研基金(20125503110012)
