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目的体外建立不同氧供状态下的细胞模型,探讨不同剂量~(60)Coγ射线对喉鳞癌Hep-2细胞周期及凋亡的影响。方法将体外培养的人喉鳞癌Hep-2细胞分为两组:A组(常氧组)、B组(低氧组),两组细胞分别接受1、3、5、10、20、40(Gy)不同剂量~(60)Coγ射线照射,两组分别设0Gy对照组。流式细胞仪检测HIF-1α蛋白表达、细胞凋亡及细胞周期变化;免疫细胞化学检测各组细胞HIF-1α蛋白表达。结果A组Hep-2细胞在接受1～5Gyγ射线照射后,主要发生了G0/G1期阻滞,在接受10～40Gyγ射线照射后,发生了G2/M期阻滞;B组细胞主要发生了G2/M期阻滞。B组0Gy对照组发生了明显的G0/G1期阻滞;A组Hep-2细胞凋亡曲线成抛物线状,凋亡率呈现明显的剂量依赖性,在5Gy处出现最大值41.56±2.25。B组细胞凋亡率同A组相比总体上呈现明显的下调趋势,但不同剂量的γ射线没有对Hep-2细胞的凋亡率产生明显的影响。结论不同剂量~(60)Coγ射线照射会导致常氧条件下Hep-2细胞发生不同周期阻滞,而不同的周期阻滞会对Hep-2细胞的凋亡产生明显的影响。低氧条件下γ射线照射未能导致Hep-2细胞发生明显的凋亡,可能同Hep-2细胞G2/M期阻滞有关,而HIF-1a蛋白的表达上调可能是Hep-2细胞G2/M期阻滞的原因之一。
OBJECTIVE: To establish a cell model of different oxygen supply states in vitro and investigate the effect of different doses of ~ (60) Coγ-rays on the cell cycle and apoptosis of Hep-2 cells. Methods Human laryngeal squamous cell carcinoma Hep-2 cells cultured in vitro were divided into two groups: group A (normoxia group) and group B (hypoxia group). The two groups of cells received 1,3,5,10,20 and 40 (Gy) different doses of ~ (60) Coγ-ray irradiation, two groups were set 0Gy control group. The expression of HIF-1αprotein, apoptosis and cell cycle were detected by flow cytometry. The expression of HIF-1αprotein was detected by immunocytochemistry. Results Hep-2 cells in group A mainly blocked G0 / G1 phase after receiving 1 ~ 5 Gy γ-ray irradiation, and G2 / M arrest occurred in 10 ~ 40 Gy γ-ray irradiation group. Cells in group B mainly occurred G2 / M block. Apoptotic curve of Hep-2 cells in group A showed obvious G0 / G1 phase arrest. The apoptosis curve of Hep-2 cells in group A was parabolic, and the apoptosis rate showed obvious dose-dependent. The maximal value was 41.56 ± 2.25 at 5 Gy. Compared with group A, the apoptotic rate of group B showed a general downward trend, but different doses of γ-rays did not significantly affect the rate of apoptosis of Hep-2 cells. Conclusion Different doses of ~ (60) Co γ -ray irradiation can lead to different cycles of Hep-2 cell arrest under normoxia, and different cycles of cell cycle arrest have a significant effect on Hep-2 cell apoptosis. Hypoxia irradiation did not lead to significant apoptosis of Hep-2 cells, which may be related to the arrest of G2 / M phase in Hep-2 cells, while the upregulation of HIF-1a protein expression may be the result of G2 / M phase block one of the reasons.