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目的:探讨乏氧诱导因子-1α(hypoxia inducible factor-1α,HIF-1α)在乏氧状态下对鼻咽癌CNE-1细胞放射敏感性的影响。方法:氯化钴处理CNE-1细胞模拟乏氧条件下的培养传代,通过脂质体将不同HIF-1α反义寡核苷酸酸导入CNE-1细胞中,根据转染复合物的不同将CNE-1细胞分为反义联合组、正义联合组、脂质体组和单纯放疗组(放疗对照组)。X射线单次照射,照射条件为照射率4 Gy/min,共8 Gy,细胞照射后继续乏氧条件下培养24 h。MTT法检测CNE-1细胞的存活率,Western blotting检测CNE-1细胞中HIF-1α和VEGF蛋白的表达。结果:在乏氧条件下,反义联合组的鼻咽癌CNE-1细胞存活率明显大于正义联合组和单纯放疗组。Western blotting结果显示,与正义联合组和单纯放疗组相比,反义联合组CNE-1细胞中HIF-1α蛋白表达显著下降(0.162±0.055 vs 0.872±0.191,0.768±0.217,0.863±0.245,P<0.05);同时反义联合组CNE-1细胞中VEGF蛋白的表达量较正义联合组和单纯放疗组明显减少(0.364±0.078 vs 1.165±0.346,1.068±0.379,1.087±0.266,P<0.05)。结论:HIF-1α反义寡核苷酸能有效抑制HIF-1α的表达,对乏氧状态下的鼻咽癌CNE-1细胞具有放射增敏作用。
Objective: To investigate the effect of hypoxia inducible factor-1α (HIF-1α) on radiosensitivity of nasopharyngeal carcinoma CNE-1 cells under hypoxic condition. METHODS: CNE-1 cells were treated with cobalt chloride to simulate culture passage in hypoxia. Different HIF-1α antisense oligonucleotides were introduced into CNE-1 cells by liposomes. According to the different transfection complex CNE-1 cells were divided into antisense group, sense combination group, liposome group and radiotherapy group (radiotherapy control group). X-ray single irradiation, the irradiation conditions 4 Gy / min irradiation rate, a total of 8 Gy, cell irradiation continue hypoxia conditions for 24 h. The survival rate of CNE-1 cells was detected by MTT assay. The protein expression of HIF-1α and VEGF in CNE-1 cells was detected by Western blotting. Results: Under hypoxic conditions, the survival rate of CNE-1 cells in antisense combined group was significantly higher than that in the combined group and radiotherapy alone group. Western blotting showed that the expression of HIF-1α in CNE-1 cells in antisense combined group was significantly lower than that in normal and radiotherapy groups (0.162 ± 0.055 vs 0.872 ± 0.191,0.768 ± 0.217,0.863 ± 0.245, P <0.05). Meanwhile, the expression of VEGF protein in CNE-1 cells in the antisense combined group was significantly lower than that in the normal and radiotherapy groups (0.364 ± 0.078 vs 1.165 ± 0.346, 1.068 ± 0.379, 1.087 ± 0.266, P <0.05) . CONCLUSION: HIF-1α antisense oligonucleotide can effectively inhibit the expression of HIF-1α and radiosensitize nasopharyngeal carcinoma CNE-1 cells under hypoxia.