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背景与目的:低氧作为实体瘤的特征和重要的生存微环境,可能与化疗耐药相关。我们认为低氧与卵巢癌化疗耐药可能相关。本研究建立低氧模型,探讨低氧、低氧诱导因子1α(HIF鄄1α)对紫杉醇诱导的人卵巢癌细胞A2780凋亡的影响。方法:体外培养的A2780细胞中加入化学性低氧诱导剂氯化钴(CoCl2),诱导并制作低氧模型。将细胞分为4组:A组(对照)、B组(常氧培养加紫杉醇)、C组(低氧培养加紫杉醇)、D组(低氧培养加Decoy加紫杉醇)。用诱骗法(Decoy)阻断HIF鄄1α功能,Westernblot、RT鄄PCR、TUNEL和流式细胞术分别检测各组细胞HIF鄄1α蛋白、mRNA的表达水平及细胞凋亡情况。结果:CoCl2能明显增加A2780细胞HIF鄄1α蛋白的表达,而对其mRNA的表达无明显影响,Decoy法阻断HIF鄄1α的功能,对其蛋白和mRNA的表达无明显影响。TUNEL检测发现,B组凋亡指数[AI:(41.12±25.65)%]显著高于C组[AI:(24.12±15.19)%](P<0.05);低氧阻断了HIF鄄1α功能后,D组凋亡指数[AI:(35.19±21.73)%]较C组明显增加(P<0.05)。流式细胞仪检测细胞凋亡率结果与TUNEL检测结果类似。结论:低氧能保护A2780细胞抵抗化学治疗,HIF鄄1α可能在紫杉醇诱导的细胞凋亡中发挥重要的抗凋亡作用。
BACKGROUND & AIM: Hypoxia, as a characteristic of solid tumors and an important living microenvironment, may be associated with chemoresistance. We think hypoxia may be related to chemoresistance in ovarian cancer. This study established hypoxia model to investigate the hypoxia, hypoxia inducible factor 1α (HIF-1α) paclitaxel-induced apoptosis in human ovarian cancer cells A2780. Methods: A2780 cells cultured in vitro were treated with cobalt chloride (CoCl2), a chemical hypoxia-inducing agent, to induce and produce hypoxia model. The cells were divided into 4 groups: group A (control), group B (normoxia plus paclitaxel), group C (hypoxia plus paclitaxel) and group D (hypoxia plus Decoy plus paclitaxel). The expression of HIF-1αprotein and mRNA were detected by Decoy, Westernblot, RT-PCR, TUNEL and flow cytometry respectively. Results: CoCl2 significantly increased the expression of HIF-1α protein in A2780 cells, but had no effect on the expression of HIF-1α. Decoy could block the function of HIF-1α and had no effect on the protein and mRNA expression. The apoptosis index of group B was significantly higher than that of group C [AI: (24.12 ± 15.19)%] (P <0.05) by TUNEL assay. After hypoxia blocked the function of HIF-1α , The apoptotic index in group D [AI: (35.19 ± 21.73)%] was significantly higher than that in group C (P <0.05). Flow cytometry apoptosis rate results and TUNEL test results similar. CONCLUSION: Hypoxia protects A2780 cells against chemotherapeutic treatment. HIF-1α may play an important anti-apoptotic role in paclitaxel-induced apoptosis.