摘要
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。
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.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2004年第2期160-162,共3页
Chinese Journal of Experimental Surgery
