低氧依赖低氧诱导因子-1α增强卵巢癌细胞SKOV3中乙酰肝素酶的表达及其侵袭力  

Hypoxia Increases the Expression of Heparitinase and the Invasiveness through the Hypoxia Inducible Factor-1α Dependent Pathway in Human Ovarian Cancer Cell Line SKOV3

作　　者：袁彩霞[1] 顾双[1] 张淑红[2] 张向宁[1] 

机构地区：[1]山东大学齐鲁医院妇产科,济南250012 [2]解放军第456医院妇产科,济南250031

出　　处：《中国医学科学院学报》2008年第6期711-716,I0010,共7页Acta Academiae Medicinae Sinicae

摘　　要：目的研究低氧条件下卵巢癌细胞株SKOV3中乙酰肝素酶(HPA)的表达及其侵袭力的变化,探讨低氧诱导因子-1α(HIF-1α)与SKOV3细胞侵袭力变化的关系。方法将SKOV3细胞分为常氧组、低氧组和低氧加雷帕霉素组。低氧组细胞在5%CO2+1%O2的条件下培养;在低氧培养前30min,向低氧加雷帕霉素组细胞培养基中加入终浓度为10ng/ml的雷帕霉素。分别于低氧培养12、24、36h收集细胞,采用Western bloting和RT-PCR方法检测HIF-1α和HPA的表达,基质凝胶侵袭实验检测细胞侵袭力。结果低氧条件下,HIF-1α的蛋白表达显著高于常氧组(P<0.05),而HIF-1αmRNA的表达不随氧浓度的降低而改变。低氧组HPAmRNA和蛋白质的表达量均显著高于常氧组(P<0.05)。雷帕霉素能抑制低氧导致的HPA mRNA和蛋白质表达的升高,HPA mRNA和蛋白质的表达均显著低于低氧组(P<0.05)。低氧组细胞的侵袭力显著高于常氧组(P<0.05)。SKOV3细胞HPA的表达与细胞侵袭力呈正相关(r=0.9863,P<0.01)。结论低氧可能依赖HIF-1α途径显著增强SKOV3细胞HPA的表达及其细胞的侵袭力。Objective To investigate the heparitinase （HPA） expression and cell invasiveness in human ovarian cancer cell line SKOV3 during hypoxia, and explore their relationship with hypoxia inducible factor-1α （HIF-1α）. Methods SKOV3 cells were incubated with normoxia, hypoxia, and hypoxia plus rapamycin, respectively. SKOV3 cells of hypoxia group were incubated in 5% CO2 ＋ 1% O2. Cells in hypoxia plus rapamycin group were incubated with 10 ng/ml of rapamycin before cultured under hypoxic condition. Cells in each group were collected for analysis after incubated with hypoxia for 12, 24, and 36 hours, respectively. Western blotting and RT-PCR were performed to detect the expressions of HIF-1α and HPA. Cell invasiveness was measured by matrigel invasion assay. Results Western blotting showed that the expression of HIF-1α significantly increased compared with normoxic group （P 〈 0. 05 ）. However, hypoxia had no obvious impact on HIF-1α mRNA expression. The expressions of HPA protein and mRNA of SKOV3 cells of hypoxia group were significantly higher than normoxic group （ P 〈 0. 05 ）. The up-regulation of HPA expression in hypoxic group decreased after the utilization of rapamycin. The cell invasion of hypoxic group was significantly higher than that of normoxic group （P 〈 0. 05 ） ; meanwhile, the expression of HPA was positively correlated with the invasiveness of SKOV3 cells （ r = 0. 9863, P 〈 0. 05 ）. Conclusion Hypoxia may promote the expression of HPA and the invasiveness of SKOV3 cells through the HIF-1α pathway, which plays an important role in the pathogenesis of ovarian cancer.

关 键 词：卵巢癌 低氧诱导因子-1Α 乙酰肝素酶 雷帕霉素 侵袭力 

分 类 号：R737.31[医药卫生—肿瘤]