外源性硫化氢通过 EGFR/PI3K/Akt 信号通路促进人卵巢癌细胞增殖、侵袭和顺铂耐药
Exogenous hydrogen sulfide promotes the proliferation, invasion and cisplatin resistance of human ovarian cancer cells through EGFR/PI3K/Akt signaling pathway

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 NaHS 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，NaHS 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 NaHS 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。
：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