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目的采用PI3K/PKB信号传导通路特异性抑制剂LY294002作用于人卵巢癌OVACA-3细胞,观察阻断PI3K/PKB信号传导通路对卵巢癌细胞增殖和凋亡等生物学特性的影响,揭示阻断PI3K/PKB信号传导通路治疗卵巢癌的机理和可行性。方法不同浓度的PI3K抑制剂LY294002作用于人卵巢癌OVACA-3细胞后,运用Western Blotting检测P-AKT及细胞核内NF-κBp65表达情况;MTT法、Annexin V-FITC流式细胞技术检测不同浓度的LY294002对卵巢癌OVACA-3细胞增殖及凋亡的影响。结果 LY294002可抑制人卵巢癌OVACA-3细胞中通路效应蛋白AKT、NF-κBp65的激活,且呈浓度及剂量依赖性。LY294002对细胞增殖抑制作用具有时间依赖性(F=10.398,P=0.001)和剂量依赖性(F=9.801,P=0.002)。LY294002可使卵巢癌细胞S期比例显著减少,G0/G1期比例增多,提示LY294002促使细胞在细胞周期G0-G1期受到阻滞。最终,抑制卵巢癌细胞增殖,促使其发生细胞凋亡,使细胞周期停滞在G1期。结论 PI3K/PKB信号传导通路阻断剂LY294002有可能成为治疗卵巢癌的新药,阻断PI3K/AKT信号传导通路可作为治疗卵巢癌的新切入点。
Objective To investigate the effects of PI3K / PKB signaling pathway-specific inhibitor LY294002 on ovarian cancer OVACA-3 cells and to observe the effects of blocking PI3K / PKB signal transduction pathway on the biological characteristics of ovarian cancer cells such as proliferation and apoptosis, Mechanism and feasibility of PI3K / PKB signal transduction pathway in treatment of ovarian cancer. Methods Different concentrations of PI3K inhibitor LY294002 were used to detect the expression of P-AKT and NF-κBp65 in human ovarian cancer OVACA-3 cells. MTT assay and Annexin V-FITC flow cytometry were used to detect the expression of P-AKT, Effect of LY294002 on proliferation and apoptosis of ovarian cancer OVACA-3 cells. Results LY294002 inhibited the activation of the effector pathways AKT and NF-κBp65 in ovarian cancer OVACA-3 cells in a dose-and dose-dependent manner. The inhibitory effect of LY294002 on cell proliferation was time-dependent (F = 10.398, P = 0.001) and dose-dependent (F = 9.801, P = 0.002). LY294002 can significantly reduce the proportion of ovarian cancer S phase, G0 / G1 phase ratio increased, suggesting that LY294002 promote cell cycle in the cell cycle G0-G1 was blocked. Finally, the inhibition of ovarian cancer cell proliferation, promote its occurrence of apoptosis, the cell cycle arrest in the G1 phase. Conclusion PI3K / PKB signaling pathway blocker LY294002 may become a new drug for the treatment of ovarian cancer. Blocking the PI3K / AKT signaling pathway may serve as a new entry point for the treatment of ovarian cancer.