雌激素受体对乳腺癌细胞软琼脂糖克隆形成及钙蛋白酶-1基因表达的影响-药理学专业论文.docxVIP

万方数据 万方数据 Effects of estrogen receptor on the gene expression of calpain1 and soft-agar colony formation in breast cancer cells. Major：pharmacology Postgraduate student：Chen Huamei Supervisors: Professor Zhu Zhuxia, Professor Wan Lei [Abstract] Objective: This study was performed to address the effect of estrogen receptor (ER) on the CANP-1 gene expression and anchorage-independent growth and the role of inhibitors for ERK/CANP in the ER-mediate action in breast cancer cells, so as to understand the mechanism that ER affects anchorage-independent growth and provide evidence for search of a more effective therapy for breast cancer. Methods: ER+/GPR30+ breast cancer cell line MCF-7 was used as a model system, 17β-estradiol(E2) and DES were applied for stimulation of model cells. Soft-agar colony formation assay was used to observe anchorage- independent growth of model cells, and RT-PCT was used to measure the levels of Capn1 mRNA . Western blot and CANP/MEK inhibitors (ALLN /U0126) were applied to observe the mediation of calpain in the above estrogen-induced action. Results: 1) ER was implicated in estrogen-induced colony formation of MCF-7 breast cancer cells. Estrogen colony formation of MCF-7 cells, increased by 55.3% （P0.05）compared to control group, while DES increased colony formation by 53.1%（P0.05）; ER antagonist ICI 182780 blocked both E2- and DES-induced action; 2) Estrogen stimulated the gene expression of calpain1 via ER. DES induced the mRNA level of calpain1 large subunit (catalytic subunit, P0.01), while that of calpain4 (regulatory subunit) was not affected significantly; 3) ER mediated colony formation and up-regulation of CANP-1 gene via ERK. MEK inhibitor U0126 pretreatment of model cells attenuated DES-induced colony formation (P0.05) and gene expression of calpain1(P0.01); 4) DES-induced colony forrnation was associated with increased calpain1 gene expression. Calpain inhibitor ALLN greatly attenuated ER-mediated calpain1 g