CAJ | 학술논문

目的：探讨雌激素（E2）对MCF-7乳腺癌细胞抗失巢凋亡能力的影响及细胞外信号调节激酶（ERK）-局部黏着斑激酶（ FAK）通路在其中的作用，以加深对E2促癌作用信号机制的认识。方法用MCF-7乳腺癌细胞，聚甲基丙烯酸羟乙基酯（ poly-Hema）涂层培养细胞诱导失巢凋亡， E2刺激及MEK和FAK抑制剂预处理细胞。用蛋白印迹法检测ERK和FAK磷酸化，台盼蓝染色细胞计数法检测细胞存活力， Hoechst荧光染色法验证细胞凋亡。结果用Poly-Hema悬浮培养可显著降低细胞存活力（ P＜0.01）， E2处理则可明显增强悬浮培养细胞的存活力（ P＜0.05），同时减少细胞凋亡；E2可引起ERK和FAK磷酸化，MEK抑制剂则可显著抑制E2诱导的ERK和FAK磷酸化，并使E2处理的悬浮培养细胞存活率下降57.48％（ P＜0.01）；FAK抑制剂能明显抑制FAK和ERK的磷酸化，同时使E2处理的悬浮培养细胞存活率下降53.59％（ P＜0.01）。结论 E2可明显提高MCF-7乳腺癌细胞对抗失巢凋亡的能力，该细胞保护效应可能与E2激活胞内ERK-FAK信号活动有关。
목적：탐토자격소（E2）대MCF-7유선암세포항실소조망능력적영향급세포외신호조절격매（ERK）-국부점착반격매（ FAK）통로재기중적작용，이가심대E2촉암작용신호궤제적인식。방법용MCF-7유선암세포，취갑기병희산간을기지（ poly-Hema）도층배양세포유도실소조망， E2자격급MEK화FAK억제제예처리세포。용단백인적법검측ERK화FAK린산화，태반람염색세포계수법검측세포존활력， Hoechst형광염색법험증세포조망。결과용Poly-Hema현부배양가현저강저세포존활력（ P＜0.01）， E2처리칙가명현증강현부배양세포적존활력（ P＜0.05），동시감소세포조망；E2가인기ERK화FAK린산화，MEK억제제칙가현저억제E2유도적ERK화FAK린산화，병사E2처리적현부배양세포존활솔하강57.48％（ P＜0.01）；FAK억제제능명현억제FAK화ERK적린산화，동시사E2처리적현부배양세포존활솔하강53.59％（ P＜0.01）。결론 E2가명현제고MCF-7유선암세포대항실소조망적능력，해세포보호효응가능여E2격활포내ERK-FAK신호활동유관。
Objective To investigate the effects of estrogen ( E2) on the resistance to anoikis and a possible role of extracellular signal-regulated kinse ( ERK)-focal adhesion kinse ( FAK) signaling in the effect of estrogen to under-stand its underlying mechanism .Methods Poly-Hema-coated culture of human breast cancer cell line MCF-7 was used to induce anoikis .Cells were treated with E2 and/or pretreated with MEK or FAK inhibitors .Western blot was used to assess the phosphorylation of ERK and FAK , trypan blue staining and cell counting were employed to evaluate cell viability , and Hoechst staining was used to check apoptosis .Results Suspension culture greatly re-duced cell survival (P<0.01), and exposure of MCF-7 cells to E2 (10 nmol/L) led to a significantly increased resistance to anoikis and survival ( P<0.05 ) as compared to DMSO .Meanwhile , E2 induced increased phospho-rylation of both ERK and FAK .Pharmacological inhibition of MEK with U 0126 ( 10 μmol/L ) reduced E2-in-creased cell survival by 57.48%(P<0.01) and E2-decreased anoikis;Treatment with FAK inhibitor (10μmol/L) attenuated E2-enhanced cell survival by 53.59% ( P<0.01 ) and E2-reduced apoptosis .Conclusions E2 con-tributes to the enhanced cell viability and increased resistance to anoikis in MCF-7 breast cancer cells , and ERK-FAK signaling may be involved in the E 2-stimulated survival during suspension culture of MCF-7 cells.