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目的:探讨表皮生长因子受体(epidermal growth factor receptor,EGFR)信号通路与卵巢癌顺铂(cisplatin,DDP)耐药细胞株SKOV3/DDP耐药相关性,及其可能的作用机制。方法:以亲本卵巢癌细胞SKOV3及DDP耐药性SKOV3/DDP细胞为研究对象;首先采用蛋白质印迹法检测EGFR信号通路在亲本株与耐药细胞中的活化情况。随后分别采用MTT法和FCM法检测单用EGFR抑制剂吉非替尼(geitinib)、DDP以及吉非替尼联合DDP对卵巢癌细胞增殖及凋亡的影响,并检测SKOV3/DDP细胞中EGFR及其下游信号通路蛋白的表达。结果:SKOV3/DDP细胞中磷酸化EGFR(phospho-EGFR,p-EGFR)(活化的EGFR)蛋白的表达量明显高于其在SKOV3细胞中的表达量(P<0.05);单独应用DDP或吉非替尼均可以抑制卵巢癌细胞的生长,吉非替尼联合DDP可明显抑制SKOV3/DDP细胞的增殖,并促进细胞凋亡,与DDP单药组相比,差异有统计学意义(P<0.05)。吉非替尼联合DDP可以明显下调p-EGFR、p-蛋白激酶B[protein kinase B,PKB(又称Akt)]和p-细胞外信号调节激酶(extracellular signal-regulated protein kinase,ERK)蛋白的表达,与DDP单药组相比,差异有统计学意义(P<0.05)。结论:吉非替尼可以增加SKOV3/DDP细胞对DDP的敏感性,其机制可能与抑制DDP诱导的EGFR及下游信号通路Akt和ERK的活性有关。
Objective: To investigate the relationship between epidermal growth factor receptor (EGFR) signaling pathway and drug resistance of cisplatin-resistant ovarian cancer cell line SKOV3 / DDP and its possible mechanism. Methods: The ovarian cancer cell line SKOV3 and DDP resistant SKOV3 / DDP cells were used as research objects. The activation of EGFR signaling pathway in parental and drug resistant cells was detected by Western blotting. Subsequently, the effects of gemtinib, DDP and gefitinib combined with DDP alone on the proliferation and apoptosis of ovarian cancer cells were detected by MTT assay and FCM assay respectively. The expressions of EGFR and Its downstream signaling pathway protein expression. Results: The expression of phospho-EGFR (p-EGFR) (activated EGFR) in SKOV3 / DDP cells was significantly higher than that in SKOV3 cells (P <0.05) FDP could inhibit the growth of ovarian cancer cells. Gefitinib combined with DDP could significantly inhibit the proliferation of SKOV3 / DDP cells and promote the apoptosis of cells. Compared with DDP group, the difference was statistically significant (P < 0.05). Gefitinib combined with DDP can downregulate the expression of p-EGFR, p-protein kinase B (PKB) and extracellular signal-regulated protein kinase (ERK) Compared with DDP group, the difference was statistically significant (P <0.05). Conclusion: Gefitinib can increase the sensitivity of SKOV3 / DDP cells to DDP, which may be related to the inhibition of DDP-induced EGFR and Akt and ERK signaling.