芪三酚与人乳腺癌MDC1基因关系的研究

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MDCl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MDCl基因与蛋白表达水平，用小RNA干扰MDCl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／LRes能明显降低MDCl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（MDCl．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MDCl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MDCl基因（MDCl-siRNA）后，MDA-MB-231细胞对Res的敏感性增加。

Effect of resveratrol on MDC1 gene expression in human breast cancer cells

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.