[1]李慧,汤小晗,卢美松*.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[J].中国计划生育和妇产科,2016,(12):15-20.
　LI Hui,TANG Xiao-han,LU Mei-song*.Study on relationship between HB-EGF and resistance of ovarian cancer to chemotherapy drug[J].Chinese Journal of Family Planning & Gynecotokology,2016,(12):15-20.
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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
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《中国计划生育和妇产科》[ISSN:1674-4020/CN:51-1708/R]

卷:
期数:
2016年12期
页码:
15-20
栏目:
论著与临床
出版日期:
2016-12-30

文章信息/Info

Title:
Study on relationship between HB-EGF and resistance of ovarian cancer to chemotherapy drug
作者:
李慧1; 汤小晗2; 卢美松*
哈尔滨医科大学附属第一医院妇产科
Author(s):
LI Hui1; TANG Xiao-han2; LU Mei-song*
Department of Gynecology and Obstetrics, The First Affiliated Hospital of Harbin Medical University, Harbin Heilongjiang 150001,P.R.China
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
Keywords:
heparin-binding epidermal growth factor-like growth factor; ovarian carcinoma; chemotherapy resistance; RNA interference; excision repair cross complementation group 1
分类号:
R 73731
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。
Abstract:
ObjectiveTo 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. MethodsIntermittent 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). Methylthiazolyl tetrazolium (MTT) assay was used to detect the cell proliferation and to calculate 50% inhibitory concentration (IC50) 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. ResultsCompared to group A, the expression level of HB-EGF proteins and IC50 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).ConclusionThe expression level of HB-EGF in cisplatin-resistant ovarian cancer cells is higher than that in sensitive cells, and inhibiting the expression of HB-EGF could enhance the sensitivity of ovarian cancer,so it could be speculated that HB-EGF is associated with resistance of ovarian cancer to chemotherapy drug. Inhibition on the expression of HB-EGF can effectively down-regulate expression of EGFR and ERCC1 proteins in cisplatin-resistant ovarian cancer cell line, which may be one of the molecular mechanisms associated with drug resistance of ovarian cancer cells.

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备注/Memo

备注/Memo:
国家自然科学基金（项目编号：81572551）
更新日期/Last Update: 2016-12-25