雌激素促进ER阴性乳腺癌细胞中IL-6的表达  

Estrogen enhances expression of interleukin-6 in ER-negative breast cancer cells

作　　者：王健[1] 徐杰[1] 安雪青 吕健东[1] 

机构地区：[1]天津市第五中心医院普外科,天津300450

出　　处：《南京医科大学学报（自然科学版）》2015年第3期309-314,共6页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家自然科学基金面上项目(81202275);天津市自然科学基金(13JCQNJC11000)

摘　　要：目的 :探讨雌激素作用下G蛋白偶联雌激素受体(G-protein coupled estrogen receptor,GPER)信号通路的活化对ER阴性乳腺癌细胞中白介素(interleukin,IL)-6表达的影响。方法:药物处理SKBR-3与MDA-MB-453细胞后,实时定量荧光PCR法检测细胞中IL-6 m RNA的变化,ELISA法检测细胞培养上清中IL-6的分泌量,Western blot法检测细胞中p-ERK与p-AKT的蛋白表达水平。结果:17-β雌二醇(E2)和GPER特异性激动剂(G1)显著促进SKBR-3与MDA-MB-453细胞中IL-6的m RNA表达以及细胞上清液中IL-6的分泌量,GPER特异性拮抗剂(G15)可显著抑制以上变化(P<0.05)。E2及G1药物处理SKBR-3细胞后显著活化细胞内GPER/AKT信号通路以及GPER/ERK信号通路(P<0.05),上调p-AKT与p-ERK的蛋白表达水平,p-AKT的相对表达量分别为对照组的(4.16±0.65)、(3.21±0.45)倍,p-ERK分别为对照组的(2.87±0.42)、(2.64±0.24)倍,在MDA-MB-453细胞中也可得到类似结果。用MEK特异性抑制剂(U0126)可明显阻断E2及G1所引发的IL-6表达变化(P<0.05),PI3K特异性抑制剂(Wortmannin)则不能。结论 :雌激素可促进ER阴性乳腺癌细胞中IL-6的m RNA表达及细胞上清液中IL-6的分泌量,其机制可能与GPER/ERK信号通路的上调有关,由GPER介导的炎症微环境可能在ER阴性乳腺癌进展中发挥重要作用。Objective：To investigate effects of activated G-protein coupled estrogen receptor（GPER） signaling pathway induced by estrogen on the production of interleukin-6 （IL-6） in ER-negative breast cancer ceils. Methods：After treatment of SKBR-3 and MDA- MB-453 cells,the expression of IL-6 mRNA was measured by Real-time qPCR. The secretion of IL-6 in supernatant was detected by ELISA. The protein expression level of p-ERK and p-AKT was determined by Western blot. Results： 17-13 estradiol （E2） and GPER specific agonist （G1） significantly increased the mRNA expression of IL-6 in SKBR-3 and MDA-MB-453 cells,which could be blocked by GPER specific antagonist （G15）. After treatment with E2 and G1, GPER/ERK and GPER/AKT signaling pathways were remarkably activated to promote the protein expression of p-ERK and p-AKT（P 〈 0.05）. The relative protein expressions of p-AKT and p-ERK in the E2 and G1 treatment groups were （4.16± 0.65）, （3.21± 0.45） and （2.87 ± 0.42）, （2.64 ± 0.24） times than those of the control group,respectively, and the same results were obtained in MDA-MB-453 ceils. Interestingly,these changes induced by E2 and G1 were significantly blocked by the MEK inhibitor U0126 rather than PI3K inhibitor Wortmannin （P 〈 0.05）. Conclusion： Estrogen enhances the expression and secretion of IL-6 in ER （-） breast cancer cells,which may associates with the up-regulation of GPER/ERK signaling pathway,and the inflammatory microenvironment mediated by GPER may play an important role in the development of ER（-） breast cancer.

关 键 词：雌激素 GPER/ERK信号通路 ER阴性乳腺癌 白介素-6 

分 类 号：R737.9[医药卫生—肿瘤]