高压氧对卵巢癌SKOV3细胞恶性生物学行为的影响及其作用机制

Effects of hyperbaric oxygen on malignant biological behavior of ovarian cancer SKOV3 cells and its related mechanism

目的探讨高压氧(hyperbaric oxygen,HBO)对卵巢癌SKOV3细胞恶性生物学行为的影响及其作用机制。方法人卵巢癌细胞系SKOV3培养完成后,HBO组和对照组分别应用CCK-8、流式细胞术、Western blotting实验检测HBO对SKOV3细胞增殖活性、凋亡及成瘤能力的影响;Western blotting检测HBO对PTEN/PI3K/AKT通路激活的影响。敲减PTEN后,再用上述方法检测HBO对SKOV3细胞增殖和凋亡的影响。采用小鼠荷瘤实验检测HBO抑制卵巢癌荷瘤小鼠肿瘤生长及相关蛋白的表达水平。结果HBO组卵巢癌细胞SKOV3增殖活性明显降低、凋亡率明显提高(均P<0.05);HBO组细胞中抑凋亡蛋白Bcl-2表达降低,而促凋亡蛋白Bax及caspase-3表达升高(均P<0.05)。HBO组明显提高了卵巢癌SKOV3细胞中PTEN蛋白的表达水平,降低了p-PI3K和p-AKT蛋白的表达水平(P<0.05或P<0.01)。此外,HBO组降低了SKOV3细胞中HIF-1的表达水平;但敲减PTEN后,HBO抑制SKOV3细胞中HIF-1表达的作用明显减弱(P<0.05),HBO+si-PTEN组SKOV3细胞增殖能力提高、凋亡受到明显抑制(均P<0.05)。与对照组相比,HBO组小鼠肿瘤体积及体质量均明显降低(均P<0.05),荷瘤卵巢癌肿瘤组织中HIF-1、p-PI3K、p-AKT、Bcl-2蛋白表达降低,而PTEN、caspase-3及Bax表达水平升高(均P<0.05)。结论HBO处理可明显抑制卵巢癌SKOV3细胞恶性生物学行为,其作用机制主要是通过调节PTEN/PI3K/AKT信号通路改善细胞缺氧环境实现的。

Objective To investigate the effect of hyperbaric oxygen(HBO)on malignant biological behavior of ovarian cancer SKOV3 cells and its related mechanism.MethodsAfter the culture of human ovarian cancer cell line SKOV3 was completed,the effects of HBO on the proliferation,apoptosis and tumorigenicity of SKOV3 cells were respectively detected by CCK-8,flow cytometry and Western blotting in the HBO group and the control group,and the effects of HBO on PTEN/PI3K/AKT pathway activation were detected by Western blotting.After PTEN was knocked down,the effects of HBO on the proliferation and apoptosis of SKOV3 cells were detected by the above methods.The inhibition of HBO on the growth of ovarian cancer-bearing mice and the expression of related proteins were detected by tumor-bearing experiment in mice.ResultsProliferation of SKOV3 cells in the HBO group was significantly decreased,while apoptosis was significantly improved(P<0.05).In the HBO group,the expression of Bcl-2 was decreased,while the expression of Bax and caspase-3 were increased(P<0.05).HBO could significantly increase the expression of PTEN protein,and at the same time decrease the expressions of p-PI3K and p-AKT proteins in ovarian cancer SKOV3 cells(P<0.05 or P<0.01).In addition,the expression of HIF-1 in SKOV3 cells was decreased in the HBO group,however,after knock-down of PTEN,the inhibitory effect of HBO on the expression of HIV-1 in SKOV3 cells was significantly weakened(P<0.05).The proliferation and apoptosis of SKOV3 cells in the hyperbaric oxygen+si-PTEN group were significantly inhibited(all P<0.05).Compared with the control group,the tumor size and body mass of mice in the HBO group were significantly decreased(all P<0.05).The expression levels of HIF-1,p-PI3K,p-AKT and Bcl-2 proteins in the ovarian cancer tissues were decreased,while the expression levels of PTEN,caspase-3 and Bax were increased(all P<0.05).ConclusionHBO treatment could significantly inhibit malignant biological behavior of ovarian cancer SKOV3 cells,the mechanism of which might be related with the improvement of the hypoxic environment of cells through the regulation of PTEN/PI3K/AKT pathway.