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目的研究MCF 7人乳腺癌细胞阿霉素敏感株 (MCF 7)和耐药株 (MCF 7/ADR)细胞中雌激素受体 (estrogenreceptor,ER)表达的变化与其对细胞生物学特性的影响。方法采用Westernblot法检测MCF 7细胞和MCF 7/ADR细胞中雌激素受体表达 ,RT PCR检测 2株细胞ERmRNA的表达水平 ,MTT法检测细胞增殖及细胞对雌激素 (estrogen ,E2 )和屈洛昔芬 (droloxifene ,Dro)的敏感性 ,流式细胞仪检测细胞周期变化。结果经Westernblot检测 ,MCF 7细胞ER为阳性 ,而MCF 7/ADR细胞ER为阴性 ,采用RT PCR可检测到MCF 7细胞中ER的mRNA ,而在MCF 7/ADR细胞则检测不到。经MTT分析 ,与MCF 7相比 ,MCF 7/ADR细胞生长速度减慢 ,细胞多分布于G0 /G1期。E2 在 1× 10 -12mol/L的浓度时开始促进MCF 7细胞的生长 ,到达 1× 10 -9mol/L时促生长作用进入平台期 ;而任何浓度的E2 对MCF 7/ADR细胞均无促生长作用。Dro的浓度达到 10 μmol/L的时候 ,开始出现对MCF 7细胞生长的浓度依赖性抑制 ,当Dro浓度达到 2 0 μmol/L时对MCF 7/ADR细胞的生长出现明显抑制 ,且高于对MCF 7细胞的抑制。结论MCF 7/ADR细胞雌激素受体表达缺失 ,缺失发生在mRNA水平以上 ,细胞生长速度减慢 ,同时细胞失去对雌激素的依赖性。
Objective To investigate the changes of estrogen receptor (ER) expression in MCF 7 cells and MCF 7 / MCF 7 cells and its effect on cell biology. Methods The expression of estrogen receptor in MCF 7 cells and MCF 7 / ADR cells was detected by Western blot. The expression of ER mRNA was detected by RT PCR. The cell proliferation and estrogen (E2) The sensitivity of droloxifene (Dro) was measured by flow cytometry. Results The results of Western blot showed that the ER of MCF 7 cells was positive and that of MCF 7 / ADR cells was negative. The mRNA of ER in MCF 7 cells was detected by RT PCR, but not in MCF 7 / ADR cells. Compared with MCF 7, the growth of MCF 7 / ADR cells slowed down and the cells distributed in G0 / G1 phase by MTT assay. E2 began to promote the growth of MCF 7 cells at the concentration of 1 × 10 -12 mol / L, and reached the plateau when the concentration reached 1 × 10 -9 mol / L; while E2 at any concentration had no effect on MCF 7 / ADR cells Growth effect. When the concentration of Dro reached 10 μmol / L, the growth inhibition of MCF 7 cells started to appear. When the concentration of Dro reached 20 μmol / L, the growth of MCF 7 / ADR cells was significantly inhibited, Inhibition of MCF 7 cells. Conclusion The expression of estrogen receptor in MCF 7 / ADR cells is absent. The deletion occurs at the level of mRNA, and the cell growth rate is slowed down. At the same time, the cells lose their dependence on estrogen.