肝素结合表皮生长因子与卵巢癌顺铂化疗耐药性关系的研究  

Study on relationship between HB-EGF and resistance of ovarian cancer to chemotherapy drug

作　　者：李慧[1] 汤小晗[1] 卢美松[1] 

机构地区：[1]哈尔滨医科大学附属第一医院妇产科,黑龙江哈尔滨150001

出　　处：《中国计划生育和妇产科》2016年第12期15-20,共6页Chinese Journal of Family Planning & Gynecotokology

基　　金：国家自然科学基金(项目编号:81572551)

摘　　要：目的检测肝素结合表皮生长因子(heparin-binding epidermal growth factor-like growth factor,HB-EGF)对卵巢癌顺铂耐药细胞增殖及耐药性的影响,探讨HB-EGF与卵巢癌顺铂耐药性的关系及机制。方法间歇大剂量冲击法诱导卵巢癌顺铂耐药细胞。使用HB-EGF siRNA转染敏感及耐药细胞,Real time PCR法检测转染效果。将实验细胞分为4组,卵巢癌敏感细胞株A 2780组(A组)、卵巢癌顺铂耐药细胞株A 2780/CDDP组(A/C组)、转染后A组(Asi组)、转染后A/C组(A/Csi组)。四甲基偶氮唑蓝比色法检测4组细胞增殖能力并计算对顺铂的半数抑制浓度(50%inhibitory concentration,IC50)。蛋白印迹法检测4组细胞中HB-EGF蛋白的表达情况,及耐药细胞转染前后表皮生长因子受体(epidermal growth factor receptor,EGFR)、核苷酸切除修复交叉互补基因1(excision repair cross complementation group 1,ERCC 1)蛋白的表达情况。结果 A/C组中HB-EGF蛋白表达量、顺铂IC50均较A组升高;与A组相比,Asi组HB-EGF蛋白表达量降低;与A/C组相比,A/Csi组中HB-EGF mRNA表达量下降,并且HB-EGF蛋白的表达量也降低,下降幅度比敏感组更大,同时A/Csi组顺铂IC50较A/C组下降;与A/C组相比,A/Csi组EGFR、ERCC 1蛋白的表达量均降低,差异均有统计学意义(P<0.05)。结论HB-EGF在卵巢癌顺铂耐药中的表达水平明显高于敏感细胞,抑制HB-EGF可以增强卵巢癌对顺铂的敏感性,提示HB-EGF与卵巢癌顺铂耐药性相关。此外,抑制HB-EGF的表达,可以下调卵巢癌顺铂耐药株中EGFR、ERCC 1蛋白的表达量,这可能是卵巢癌顺铂耐药的相关机制之一。Objective To explore the effects of heparin- binding epidermal growth factor- like growth factor （HB -EGF） on proliferation and drug - resistance of cisplatin - resistant ovarian carcinoma cells, to discuss the relationship between HB - EGF and cisplatin resistance of ovarian carcinoma and its mechanism. Methods Intermittent high - dose shocking method induced cisplatin -resistant ovarian cancer cells. HB - EGF siRNA was used to transfect sensitive and drug - resistant cells, and real - time - PCR was used to detect transfection effect. Test cell was divided into four groups, sensitive ovarian cancer cell line A2780 group （A group）, cisplatin -resistant ovarian cancer cell line A2780 / CDDP group （A / C group） , group A after transfection （Asi group） , A/C group after transfection （A/Csi group）. Methyhhiazolyl tetrazolium （MTr） assay was used to detect the cell proliferation and to calculate 50% inhibitory concentration （ICS0） of 4 groups of cells to cisplatin. Western blot was used to detect expressions of HB - EGF protein in 4 groups of cells and the expressions of epidermal growth factor receptor （EGFR） and excision repair cross complementation group 1 （ ERCC1 ） in cells of drug - resistance groups before and after transfection. Results Compared to group A, the expression level of HB - EGF proteins and ICS0 of group A/C was higher ; Compared to group A, the expression of HB - EGF proteins in group Asi was decreased. Compared to group A/C, the expression level of HB - EGF mRNA of group A/Csi was decreased, and the expression of HB -EGF proteins in group A/C wase significantly decreased, the decline of group A/C is more obvious;at the same time, the IC50 of group A/Csi was decreased compared with group A/C. Compared to group A/C, the expression of EGFR and ERCC1 proteins were both reduced after inhibition on HB - EGF expression （ P 〈 0. 05 ）. Conclusion The expression level of HB - EGF in cisplatin - resistant ovarian cancer cells is higher than that in sensitive cells,

关 键 词：肝素结合表皮生长因子 卵巢癌 化疗耐药性 RNA干扰 核苷酸切除修复交叉互补基因 

分 类 号：R737.31[医药卫生—肿瘤]