目的

将雌激素受体ERβ1 真核表达质粒转染到人乳腺癌MDA-MB-231细胞中,观察外源性ERβ1 基因转染MDA-MB-231 细胞后对E-cadherin、Vimentin 等基因表达和细胞上皮间质转化的影响,探讨ERβ1 在乳腺癌发生发展中的生物学作用机制。

方法

应用脂质体法将ERβ1 真核表达质粒转染至乳腺癌MDA-MB-231 细胞中,分别用实时荧光定量PCR、Western Blot 检测转染前后细胞中ERβ1、E-cadherin、Vimentin mRNA 和蛋白表达的变化;并用细胞增殖曲线显示转染后细胞增殖能力的改变。 所有数据以 表示,多个样本均数间的比较用单因素方差分析,组间比较采用SNK 法。 细胞生长曲线变化采用重复测量方差分析。

结果

外源性ERβ1 真核表达质粒转染组MDA-MB-231 细胞较未转染组ERβ1、E-cadherin mRNA 和蛋白水平明显增强(P<0.010),Vimentin mRNA 水平明显减少(P<0.010),细胞增殖能力明显减弱。

结论

ERβ1 可能参与抑制乳腺癌MDA-MB-231 细胞的上皮间质转化过程。

Objective

To explore the effect of exogenous ERβ1 gene on the expression of E-cadherin and vimentin by transfecting recombinant eukaryotic expressing vector containing ERβ1 cDNA into human breast cancer MDA-MB-231 cells, and to investigate the biological role of ERβ1 in epithelial-mesenchymal transition of breast cancer cells.

Methods

Recombinant eukaryotic expressing vector containing ERβ1 cDNA was transfected into human breast cancer MDA-MB-231 cells. The mRNA and protein expression levels of ERβ1, E-cadherin and vimentin were tested by real-time polymerase chain reaction and Western blot, respectively. The cell growth curve showed the change of proliferation ability in MDA-MB-231. All data were expressed as . Single factor analysis of variance was used to compare the multiple sample means, and SNK method for group comparison.Changes in the cell growth curve were analyzed using repeated ANOVA.

Results

The mRNA and protein expression levels of ERβ1 and E-cadherin increased significantly in the transfection group compared with the control group (P<0.010). The vimentin mRNA level decreased significantly in the transfection group compared with the control group(P<0.010).The proliferation ability of the transfection group decreased either.

Conclusion

ERβ1 may play a role in inhibiting the epithelial-mesenchymal transition of breast cancer MDA-MB-231 cells.