论文部分内容阅读
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 investigate the effect of heparin-binding epidermal growth factor-like growth factor (hf-EGF) on the proliferation and drug resistance of cisplatin-resistant ovarian cancer cells and explore the relationship between HB-EGF and cisplatin-resistant ovarian cancer Sexual relations and mechanisms. Methods Intermittent high dose shock induced cisplatin - resistant ovarian cancer cells. Transfection of sensitive and resistant cells with HB-EGF siRNA and Real time PCR assay of transfection efficiency. The experimental cells were divided into four groups: A 2780 group (A group), A 2780 / CDDP group (A / C group), A group (Asi group) , A / C group (A / Csi group) after transfection. The proliferation of 4 groups of cells was detected by MTT assay and the 50% inhibitory concentration (IC50) of cisplatin was calculated. The expression of HB-EGF protein in four groups of cells was detected by Western blotting, and the expression of EGFR, excision repair cross complementation group 1, ERCC 1) protein expression. Results Compared with group A, the expression of HB-EGF protein and the IC50 of cisplatin in group A / C were higher than those in group A; The expression of HB-EGF in group A was lower than that in group A; Compared with group A / C, The expression of HB-EGF mRNA was decreased and the expression of HB-EGF protein was also decreased in group A and C than in A / C group, while the IC50 of A / Csi group was lower than that of A / C group The expression of EGFR and ERCC1 protein in A / Csi group was lower than that in A / Csi group (P <0.05). Conclusion The expression of HB-EGF in cisplatin-resistant ovarian cancer cells was significantly higher than that of the sensitive cells. Inhibition of HB-EGF could enhance the sensitivity of ovarian cancer to cisplatin, suggesting that HB-EGF is associated with cisplatin resistance in ovarian cancer. In addition, inhibiting the expression of HB-EGF can down-regulate the expression of EGFR and ERCC-1 in cisplatin-resistant ovarian cancer cells, which may be one of the mechanisms of cisplatin-resistant ovarian cancer.