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目的:探讨了乳腺癌细胞中膜雌激素受体(membraneestrogenreceptor,m ER)表达情况以及对表皮生长因子水平的影响。方法:采用q RT-PCR和Western blot分析乳腺癌细胞乳腺癌细胞系-7(michigan cancer foundation-7,MCF-7)、MDA-MB-231和MDA-MB-435中膜雌激素受体m ER m RNA和蛋白质的表达水平。并选取中等表达的细胞采用慢病毒介导的m ER sh RNA建立m ER Knowdown的稳定细胞系,并分析膜雌激素白蛋白-E2(membrane estrogen-2,m E2)和传统的雌激素(estrogen-2,E2)分别对乳腺癌细胞(ERα+和ERα-)的生长影响及其对EGFR和EGF表达水平的影响。结果:3种乳腺癌细胞系中m ER均有表达,其中MCF-7细胞表达水平最高,MDA-MB-231和MDA-MB-435细胞表达水平相当。采用慢病毒介导m ER sh RNA建立的MCF-7稳定细胞系(MCF-7/ERα-)其m ER m RNA和蛋白质水平较正常MCF-7均明显下调(P<0.05)。传统E2均能促进MCF-7和MCF-7/ERα-细胞的增殖,无统计学意义(P>0.05),但m E2作用后,MCF-7/ERα-生长速度明显低于MCF-7细胞(P<0.05)。传统E2刺激MCF-7和MCF-7/ERα-细胞后,细胞EGF m RNA和蛋白质水平无差异(P>0.05),而m E2刺激MCF-7和MCF-7/ERα-细胞后,MCF-7/ERα-细胞EGF m RNA和蛋白质水平较MCF-7细胞水平明显下降(P<0.05)。结论:本研究有望为乳腺癌的治疗提供更新的分子治疗靶点,更精确的临床内分泌治疗预测指标。
Objective: To investigate the expression of membrane estrogen receptor (m ER) in breast cancer cells and its effect on the level of epidermal growth factor. Methods: RT-PCR and Western blot were used to detect the expression of estrogen receptor m in breast cancer cell line-7 (MCF-7), MDA-MB-231 and MDA-MB-435 ER m RNA and protein expression levels. The moderately expressed cells were used to construct m ERKnowdown stable cell line using lentivirus-mediated m ER-sh RNA. The effects of membrane estrogen-2 (E2) and traditional estrogen -2, E2) on the growth of breast cancer cells (ERα + and ERα-) and their effects on the expression of EGFR and EGF, respectively. Results: The expression of m ER was found in all the three breast cancer cell lines, of which MCF-7 cells had the highest level of expression and MDA-MB-231 and MDA-MB-435 cells had the same level of expression. MCF-7 stable cell line (MCF-7 / ERα-) established by lentivirus-mediated m ER sh RNA had significantly lower m ER m RNA and protein levels than normal MCF-7 cells (P <0.05). Traditional E2 could promote the proliferation of MCF-7 and MCF-7 / ERα-cells, but there was no statistical significance (P> 0.05). However, the growth rate of MCF-7 / ERα- (P <0.05). The expression of EGF mRNA and protein in MCF-7 and MCF-7 / ERα-cells was not significantly different after stimulated by traditional E2 (P> 0.05), while MCF-7 and MCF-7 / ERα- 7 / ERα-cells EGF m RNA and protein levels were significantly decreased compared with MCF-7 cells (P <0.05). Conclusion: This study is expected to provide new therapeutic targets for molecular breast cancer therapy, more accurate prediction of clinical endocrine therapy.