知识与财富的链接
目的将雌激素受体(ER)β1真核表达质粒转染人乳腺癌MDA-MB-231细胞中,观察外源性ERβ1基因转染MDA-MB-231细胞后对p21基因表达和细胞增殖能力的影响,探讨ERβ1在乳腺癌中的生物学作用机制。方法应用脂质体法将ERβ1真核表达质粒转染至乳腺癌MDA-MB-231细胞中,流式细胞仪观察细胞凋亡率的变化;分别用实时聚合酶连锁反应(RT-PCR)、Western Blot检测转染前后细胞中ERβ1、p21mRNA和蛋白表达的变化;细胞增殖曲线显示转染后细胞增殖能力的改变。统计分析采用t检验和单因素方差分析。结果外源性ERβ1真核表达质粒转染组MDA-MB-231细胞较未转染组ERβ1、p21mRNA和p21蛋白水平明显增强(P0.010),细胞增殖能力明显减弱,细胞凋亡率从1.4%升至6.14%(t=-7.960,P=0.001)。结论 ERβ1可以通过上调p21基因抑制乳腺癌MDA-MB-231细胞的增殖。
Objective To explore the effect of exogenous estrogen receptor-β1（ERβ1）gene on the expression of p21as well as the changes in proliferation ability of MDA-MB-231cell line by transfecting recombinant eukaryotic expressing vector containing ERβ1cDNA into human breast cancer cell MDA-MB-231,and to investigate the biological role of Erβ1in breast cancer.Methods Recombinant eukaryotic expressing vector containing ERβ1cDNA was transfected into human breast cancer cell MDA-MB-231by using cationic liposome Lipofectamine 2000TMas transfecting agent.The expression levels in both mRNAs and the protein levels of both ERβ1and p21 were tested by real-time polymerase chain reaction（RT-PCR）and Western blot,respectively.The changes of proliferation ability in MDA-MB-231was displayed by cell growth curve,and the changes in cell apoptosis were detected by flow cytometry.Statistical analysis was performed using t test and analysis of variance.Results The proliferation ability decreased and the expression levels of both ERβ1and p21mRNA and protein increased significantly in the transfection group compared with the control group（P〈0.010）.The rate of cell apoptosis increased from 1.4%to 6.14%（t=-7.960,P=0.001）.Conclusions ERβ1can inhibit the cell growth of MDA-MB-231 cells by upregulating the expression of p21.