摘要
目的·探讨外源性硫化氢对人卵巢癌细胞增殖、凋亡、侵袭和顺铂耐药的作用及其机制。方法·以人卵巢癌细胞株SKOV3细胞和人顺铂耐药卵巢癌细胞株SKOV3/DDP细胞为研究对象。采用CCK-8法、流式细胞术、Transwell侵袭小室实验,分别检测NaHS对SKOV3细胞的增殖、凋亡、侵袭能力的影响;通过计算细胞半数抑制浓度(half maximal inhibitory concentration,IC50)和细胞活性抑制率(inhibition rate,IR),检测NaHS对SKOV3和SKOV3/DDP细胞顺铂耐药的影响;Western blotting检测Na HS对SKOV3和SKOV3/DDP细胞中EGFR、PI3K和Akt磷酸化水平的影响;用EGFR的抑制剂erlotinib、PI3K的抑制剂LY294002和Akt的抑制剂MK-2206处理细胞,检测SKOV3和SKOV3/DDP细胞中EGFR、PI3K和Akt的磷酸化水平以及SKOV3细胞增殖、侵袭和SKOV3/DDP细胞顺铂耐药的变化。结果·与对照组相比,Na HS能显著促进SKOV3细胞增殖(P=0.000)和侵袭(P=0.033);显著提高SKOV3和SKOV3/DDP细胞对顺铂的IC50(P=0.027,P=0.009),降低顺铂对细胞的IR(P=0.001,P=0.009);激活SKOV3和SKOV3/DDP细胞的EGFR(P=0.000,P=0.037)、PI3K(P=0.009,P=0.013)、Akt(P=0.000,P=0.023);erlotinib、LY294002和MK-2206均能阻断Na HS对SKOV3细胞的增殖(均P=0.000)和侵袭(均P<0.01)作用,也能逆转NaHS促进SKOV3/DDP细胞对顺铂耐药的作用(均P=0.000)。结论·外源性硫化氢可促进SKOV3细胞的增殖和侵袭,以及SKOV3和SKOV3/DDP细胞的顺铂耐药,其机制与激活EGFR/PI3K/Akt信号通路有关。
Objective·To investigate the effects and mechanisms of exogenous hydrogen sulfide on the proliferation,apoptosis,invasion and cisplatin resistance of human ovarian cancer cells.Methods·The human ovarian cancer cell line SKOV3 cells and human cisplatin resistant cell line SKOV3/DDP cells were studied.The effects of NaHS on cell proliferation,apoptosis and invasion in SKOV3 cells were detected respectively by CCK-8,flow cytometry and Transwell invasion assay.The effect of NaHS on cisplatin resistance in SKOV3 and SKOV3/DDP cells was detected by calculating the IC50 and IR.The phosphorylation levels of EGFR,PI3K and Akt in SKOV3 and SKOV3/DDP cells were assayed by Western blotting.After treated with erlotinib(EGFR inhibitor),LY294002(PI3K inhibitor)and MK-2206(Akt inhibitor),the phosphorylation levels of EGFR,PI3K and Akt in SKOV3 and SKOV3/DDP cells,as well as cell proliferation,invasion in SKOV3 cells and cisplatin resistance in SKOV3/DDP cells were detected.Results·Compared with the control group,NaHS could significantly promote the proliferation(P=0.000)and invasion(P=0.033)in SKOV3 cells;increase IC50(P=0.027,P=0.009)and decrease IR of cisplatin(P=0.001,P=0.009)in SKOV3 and SKOV3/DDP cells.NaHS could activate EGFR(P=0.000,P=0.037),PI3K(P=0.009,P=0.013)and Akt(P=0.000,P=0.023)in SKOV3 and SKOV3/DDP cells.Erlotinib,LY294002 and MK-2206 could block the effects of NaHS on the proliferation(all P=0.000)and invasion(all P<0.01)in SKOV3 cells,and also reverse the effect of NaHS on the cisplatin resistance in SKOV3/DDP cells(all P=0.000).Conclusion·Exogenous hydrogen sulfide can induce the proliferation and invasion in SKOV3 cells,and promote the cisplatin resistance in SKOV3 and SKOV3/DDP cells,which mechanisms are related to activation of EGFR/PI3K/Akt signaling pathway.
作者
葛顺娜
段峥峥
GE Shun-na;DUAN Zheng-zheng(Institute of Embryo-Fetal Original Adult Disease,International Peace Maternity&Child Health Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200030,China)
出处
《上海交通大学学报（医学版）》
CAS
CSCD
北大核心
2018年第3期244-253,共10页
Journal of Shanghai Jiao tong University:Medical Science
基金
国家自然科学基金(81601335)
上海市自然科学基金(15ZR1444000)
