雌激素受体真核表达质粒对乳腺癌耐药细胞耐药性和细胞增殖的影响

目的　研究雌激素受体 (ER )表达质粒对MCF 7/ADR乳腺癌阿霉素耐药细胞耐药性和细胞增殖的影响。方法　Westernblot法检测MCF 7细胞和MCF 7/ADR细胞ER状态。构建ER的真核细胞表达质粒 pCER ,导入MCF 7/ADR细胞 ,流式细胞仪检测细胞周期分布 ,MTT法检测细胞生长。结果　MCF 7细胞ER为阳性 ,MCF 7/ADR细胞ER为阴性。成功构建真核细胞表达质粒 pCER ,转染MCF 7/ADR细胞 ,获得阳性克隆MTER/ADR。与对照组相比 ,MTER/ADR生长速度增快 ,S期细胞为 17.2 3 % (对照组为 14 .66% ) ,细胞对阿霉素的耐药性下降 ,阿霉素为 3mg/L时对细胞的抑制率为 45 % ,大于对照组细胞的抑制率 (为 2 4% ) ;且雌二醇 (E2 )能增加阿霉素对MTER/ADR细胞生长的抑制作用。结论　MCF 7/ADR乳腺癌耐药细胞的耐药性与其雌激素受体的表达缺失有一定关系。

Effects of estrogen receptor expression plasmid on the drug resistance and growth of drug-resistant human breast cancer cells

Objective To study the effect of ER expression plasmid on drug resistance and growth of drug-resistant MCF-7/Adr human breast cancer cells.Methods ER status of MCF-7 cells and MCF-7/ADR cells was analyzed by Western blot.ER expression plasmid was constructed and transfected into MCF-7/ADR cells via LipofectAMINETM.After G418 screening,the positive clone MTER/ADR was obtained.The integration and expression of ER gene were detected by PCR and Western blot.The growth and drug resistance to Adriamycin of MTER/ADR cells were investigated by MTT assay.The cell cycle of MTER/ADR cells was monitored by flow cytometry.Results ER was positive in MCF-7 cells,but was negative in MCF-7/ADR cells.The ER expression plasmid was constructed and transfected into MCF-7/ADR cells via LipofectAMINETM.The integration and expression of ER gene were successful.The MTER/ADR cells showed faster growth rate and lower drug resistance to Adriamycin (the inhibition rate was 45 % at the concentration of 3 mg/L while the control was 24 %),and the cell cycle of MTER/ADR cells was arrested at S phase (the percent of S phase was 17.23 % while the control was 14.66 %).Conclusion The drug resistance of MCF-7/ADR cells was associated with the ER status.Estrogen could stimulate the growth and reverse the drug resistance to Adriamycin of MCF-7/ADR cells via ER expression plasmid.