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目的 检测miyabenolC(MiyC)和kobophenolA(KobA)是否通过依赖雌激素受体 (ER)发挥促雌激素受体阳性细胞MCF 7生长的作用。方法 构建雌激素受体反义表达质粒 (pCASER) ,转染MCF 7细胞株 ,经G418筛选 ,PCR、Westernblot鉴定筛选出阳性表达克隆 (MTASER)。运用MTT法检测药物对细胞生长状况的影响。运用放射竞争结合实验检测药物与ER的结合。结果 MiyC和KobA对MCF 7细胞的促生长作用大于对MTASER细胞的作用 ;MiyC和KobA均可竞争性抑制ER与 17β 雌二醇的结合。 结论 在一定浓度范围内 ,MiyC和KobA依赖ER发挥促细胞生长的作用。
Objective To determine whether miyabenolC (MiyC) and kobophenolA (KobA) play a role in the growth of estrogen-receptor positive cells MCF-7 through ER-dependent estrogen receptor (ER). Methods The estrogen receptor antisense plasmid (pCASER) was constructed and transfected into MCF - 7 cells. After screening by G418, the positive expression clones (MTASER) were screened by PCR and Western blot. The effects of drugs on cell growth were detected by MTT assay. Radiation competition was used to test the combination of drug and ER. Results MiyC and KobA promoted the growth of MCF 7 cells more than MTASER cells. Both MiyC and KobA could competitively inhibit the binding of ER to 17β estradiol. Conclusion MiyC and KobA depend on ER to promote cell growth in a certain concentration range.