雌激素通过细胞外信号调节激酶-钙蛋白酶-局部黏着斑激酶通路抑制乳腺癌细胞失巢凋亡-病理学与病理生理学专业论文.docxVIP

贵阳医学院 贵阳医学院 2015 届科学学位硕士研究生学位论文 万方数据 万方数据 激素能够明显提高乳腺癌 MCF-7 细胞的抗失巢凋亡能力； 2) 雌激素的抗失巢 凋亡效应与激活胞内 ERK/CANP/FAK 信号通路活动有关; 3) 乳腺癌细胞 CANP 活性增高及 CANP/FAK 与 ERK 之间的正反馈环路可能增强乳腺癌细胞对雌激素 刺激的反应性。 [关键词] 雌激素; 细胞外信号调节激酶; 局部黏着斑激酶; 钙激活中性蛋白 酶;失巢凋亡; 乳腺癌细胞 III Estrogen suppresses anoikis via extrcellular signal-regualted kinase (ERK)-calpain-focal adhesion kinase (FAK) signaling in breast cancer cells2 Major: PathophysiologyPostgraduate student: He YanSupervisors: Prof. Wang Xudong; Prof. Yang Qin [Abstract] Objective: This study was performed to investigate the effects of estrogen（E2）on the resistance to anoikis and a possible role of calpain-focal adhesion kinse (FAK) signaling underlying the estrogen action in breast cancer cells, for the purpose of searching a high-efficiency therapy. Methods: Human breast cancer cell line MCF-7(ER+) was employed as a model system and poly-Hema-coated culture was used to induce anoikis and where required, estrogen-sensitive MCF-10A and estrogen insensitive- MDA-MB-231 cells were used. Cells were treated with E2 and/or pretreated with MEK or FAK inhibitors where needed. Western blotting was used to assess the phosphorylation of ERK and FAK, MTT assay and trypan blue staining/ cell counting were employed to evaluate cell viability, and Hoechst staining was used to verify apoptosis. Results: 1) E2 enhanced anoikis resistance and increased phosphorylation of ERK and FAK in MCF-7 breast cancer cells. Compared to anchorage-dependent culture, suspension culture with poly-hema profoundly reduced survival ； Treatment of model cells in suspension culture dramatically decreased cell survival and apoptosis. Additionally, E2 treatment led to increased phosphorylation of ERK and FAK. 2) Inhibitors for ERK or calpain greatly reversed phosphorylation of FAK and anti-anoikis effect induced by E2 stimulation. Treatment of MCF-7 cells with E2 for 12 h significantly enhanced phosphorylation of ERK and FAK, while MEK inhibitor U0126 treatment suppress