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目的探讨雌激素受体β亚型(ERβ)对人乳腺癌细胞株生物学特性的影响和作用机制。方法构建ERβ稳定高表达的MDAMB435细胞株。运用噻唑蓝(MTT)法、流式细胞术和Transwell等方法,观察不同雌激素浓度下ERβ对该细胞株增殖、侵袭和转移特性的影响。采用RTPCR和(或)Westernblot、明胶酶谱技术检测相关基因的表达水平。结果ERβ可显著提高MDAMB435细胞的增殖速度和侵袭迁徙能力,且呈非雌激素依赖性。与对照细胞相比,ERβ高表达的细胞S期比例显著增多(P=0.01);p21mRNA表达水平降低33.3%(P=0.03),蛋白表达水平降低47.4%(P=0.02);基质金属蛋白酶(MMP)9mRNA表达水平增高91.3%(P<0.01),活性增高67.3%(P=0.02);Ets1mRNA表达水平增高62.2%(P=0.01),蛋白表达水平增高51.0%(P=0.01);cyclinA、cyclinE、cyclinD1、MMP1、MMP2、MMP7、血管内皮生长因子(VEGF)和碱性成纤维细胞生长因子(bFGF)在mRNA水平未见明显差异。结论ERβ有促进乳腺癌细胞增殖、侵袭和转移的作用。降低p21的表达、增加Ets1和MMP9的表达并增强MMP9的活性是其可能的作用机制之一。
Objective To investigate the effect and mechanism of estrogen receptor beta subtype (ERβ) on the biological characteristics of human breast cancer cell lines. Methods MDAMB435 cell line with stable expression of ERβ was constructed. The effect of estrogen on the proliferation, invasion and metastasis of the cell line was observed by MTT assay, flow cytometry and Transwell assay. RTPCR and / or Western blot were used to detect the expression of related genes. Results ERβ significantly increased the proliferation, migration and invasion of MDAMB435 cells in a non-estrogen-dependent manner. Compared with the control cells, the proportion of S phase in ERβ-overexpressed cells increased significantly (P = 0.01); the expression of p21mRNA decreased by 33.3% (P = 0.03) and the protein expression decreased by 47.4% (P = 0.02) (P = 0.01). The expression of cyclin A, MMP 9 mRNA increased by 91.3% (P <0.01), the activity increased by 67.3% (P = 0.02); the expression of Ets1 mRNA increased by 62.2% CyclinE, cyclinD1, MMP1, MMP2, MMP7, VEGF and bFGF had no significant difference at mRNA level. Conclusion ERβ can promote the proliferation, invasion and metastasis of breast cancer cells. Reducing the expression of p21, increasing the expression of Ets1 and MMP9 and enhancing the activity of MMP9 are one of its possible mechanisms.