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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 on the inhibition of proliferation of human breast cancer MDA-MB-231 cells and its relationship with MDC1 gene. Methods The human breast cancer MDA-MB-231 cell line was used as the research object. The cell proliferation was determined by MTS method. The effect of Res on breast cancer MDA-MB-231 cells was observed by acridine orange fluorescence staining. Methods The MDC1 gene and protein expression levels were determined. After MDC1 gene was silenced by small RNA, the apoptosis of MDC1 cells was detected by flow cytometry and the sensitivity to Res was observed. Results Res (40μmol / L) significantly inhibited the proliferation of breast cancer MDA-MB-231 cells (P <0.05), while 0,60 and 120μmol / L Res significantly decreased the expression of MDC1 gene and protein (P <0.05). Flow cytometry analysis showed that the apoptosis rate of MDC1-siRNA group was (45.13 ± 6.2)% compared with that of the negative control group [(24.34 ± 2.6)%] and the untreated group Group [(17.69 ± 4.9)%] increased significantly (P <0.05). MTS results showed that the sensitivity of cells to Res increased after MDC1 gene interference. Conclusion Res above 40μmol / L can inhibit the proliferation of MDA-MB-231 cells. Res can effectively reduce the expression of MDC gene and protein and promote the apoptosis of MDA-MB-231 cells. The sensitivity of MDA-MB-231 cells to Res increased after the small RNA interference with the MDC1 gene (MDC1-siRNA).