缺氧状态下人胰腺癌细胞对吉西他滨反应的实验研究

Hypoxia increase the resistance of human pancreatic cancer cells to gemcitabine chemotherapy

目的:观察缺氧状态对人胰腺癌细胞吉西他滨化疗敏感性的影响并分析其相关机制。方法:将人胰腺癌细胞SW1990分为常氧组、缺氧组、常氧+吉西他滨组和缺氧+吉西他滨组。MMT法检测各组细胞增殖、流式细胞仪检测细胞凋亡、Real-time PCR和Western blot分别检测缺氧诱导因子-1α(HIF-1α)、多药耐药基因(MDR-1)mRNA和蛋白的表达。结果:与常氧+吉西他滨组相比,低氧+吉西他滨组的细胞增殖率明显增加,细胞凋亡率显著下降(P<0.05);低氧组和低氧+吉西他滨组HIF-1α蛋白表达分别显著高于常氧组(P<0.05或P<0.01);与常氧组相比,低氧组和低氧+吉西他滨组的MDR1 mRNA和蛋白表达水平均显著升高(P<0.05或P<0.01)。结论:缺氧状态可增加人胰腺癌细胞SW1990对吉西他滨的化疗抵抗,其机制与缺氧环境可诱导HIF-1α和MDR1基因表达有关。

Objective：To observe the impact of the hypoxic state of human pancreatic carcinoma cell on gemcitabine chemosensitivity and the mechanism. Methods： SW1990 pancreatic cancer ceils were divided into normoxic group,hypoxia group, normoxic plus gemcitahine group, and hypoxia plus gemcitabine group. Cell proliferation were detected by MMT and apoptosis were detected by flow cytometry. Real -time PCR and Western blot were used to detect hypoxia - inducible factor - 1 α （ HIF - 1 α ） , the multidrug resistance gene （ MDR - 1 ） mRNA and protein expression. Results： The proliferation rate of hypoxia ＋ gemcitabine group compared to the normoxia plus gemcitabine group was significantly increased and the apoptotic rate was significantly decreased （P 〈 0.05 ）. H/F - 1α protein expression in hypoxia group and hypoxia ＋ gemcitabine group were significantly higher than the normoxic group （ P 〈 0.05 or P 〈 0.01 ）. Compared with normoxic group, MDR1 mRNA and protein significantly increased in hypoxie group and hypoxia ＋ gemcitabine group （P 〈0.05 or P 〈 0.01 ）. Conclusion： Hypoxia can increase chemotherapy resistance to gemcitabine of SW1990 pancreatic cancer cell, and it was caused by HIF - Ice and MDR1 gene expression induced by hypoxic environment.