【摘要】 目的通过脂质体转染技术,将ERβ转染给ERα+的MCF-7细胞和ERα-的MDA-MB-231细胞,研究在ERα+和ERα-的不同条件下ERβ的功能。方法利用脂质体转染试剂Lipofectmine LTX将质粒pReveicer-M72-ERβ对MCF-7细胞和MDA-MB-231进行转染,并将细胞分为MCF-7组(ERα+ERβ-)、MCF-7ERβ组(ERα+ERβ+)、MCF-7空组(ERα+ERβ-)、MDA-MB-231组(ERα-ERβ-)、MDA-MB-231 ERβ组(ERα-ERβ+)、MDA-MB-231空组(ERα-ERβ-),对各组细胞进行雌二醇、他莫昔芬处理,并同时设置未处理组作为对照。用MTT法绘制各组细胞在不同处理因子作用下的生长曲线。结果①与MDA-MB-231组和MDA-MB-231空组相比,MDA-MB-231 ERβ组细胞生长速度加快(P<0.05),三组细胞对于雌二醇、他莫昔芬的反应差异无显著性;②与MCF-7组和MCF-7空组对比,MCF-7ERβ组细胞生长速度减慢(P<0.05);在雌二醇的作用下,MCF-7ERβ组细胞的生长速度比其他两组细胞减慢(P<0.05),三组细胞对于他莫昔芬的反应差异无显著性。结论①在ERα表达的条件下,ERβ的表达能够抑制细胞的生长,并且能够使细胞对于雌激素的敏感性下降;②在无ERα表达的条件下,ERβ的表达能够促进细胞的生长。更多还原

【Abstract】 【Objective】To study the effects of exogenous ERβ on the growth of breast cancer MCF-7 and MDA- MB-231 cells under different treatment with estrogen or tamoxifen; to discuss ERβ’s biological function and the re- lation between ERα and ERβ; to explore the possibility that ERβ can be used to predict the reaction of endocrine therapy together with ERα. 【Methods】①Extracted, tested and verified plasmid pReveicer-M72-ERβ which con- tains ER β gene through agarose gel electrophoresis experiment. ②An eukaryotic expression vector containing 1.6 kb of human entire coding sequence of ERβ(pReveicer-M72-ERβ) was transfected into MCF-7 and MDA-MB-231 cells using Lipofectmine LTX, empty vector group and blank control group were used to two kinds of cells. ③20 hours later after transfection, fluorescence microscope,immunefluorescent, Western Blot were used to validate ER β expression. ④For six groups of MCF-7, MCF-7ERβ, MCF-7, MDA-MB-231, MDA-MB-231 ERβ, MDA-MB-231, the growth properties of each group under different treatment, including E2 and tamoxifenwere observed using MTT and untreated group as control.【Results】①MDA-MB-231 ERβ group cells grow faster than MDA-MB-231 and MDA-MB-231, but there was no difference among three group cells under E2 and tamoxifen. ②ERβ expression in MCF-7ERβ inhibited cell growth,and inhibition of proliferation of MCF-7ERβ in the presence of E2 was observed, MCF-7ERβ cells proliferated at the same rate as MCF-7 and MCF-7 in the presence of tamoxifen.【Conclusions】 ①In the presence of ERα, expression of ERβ inhibited cell growth, reduced the sensitivity of the cells to estrogen, ②When there was no ERα, expression of ERβ can accelerate cell growth, but have no influence on cells under the treatment of estrogen and tamoxifen.更多还原