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目的研究p53基因在电离辐射诱导的非小细胞肺癌细胞死亡中的作用。方法选取H1299(p53-/-),H1299-P53,H1299-175H细胞为研究对象。采用流式细胞术检测细胞凋亡率,WESTERN BOLT方法检测相关蛋白表达情况。结果在H1299(P53-/-)细胞中,4GY、8GY照射后细胞凋亡率分别是假照组的68.4%、50%。在H1299-P53细胞中,4GY、8GY照射后细胞凋亡率分别是假照组的1.7倍、8.1倍。在H1299-175H细胞中,4GY8、GY照射后细胞凋亡率分别是假照组的1.5倍、2倍。LC3蛋白表达在H1299呈现剂量依赖性升高,在H1299-175H呈现剂量依赖性降低,在H1299-P53中,OGY表达最高,4 GY表达最低,8 GY表达低于假照组但比4 GY高。AKT表达在H1299细胞呈剂量依赖性降低,H1299-P53在4GY表达量升高而在8GY又降低但仍高于假照组,在H1299-175H细胞中AKT表达呈剂量依赖性降低。MDM2在H1299的表达呈剂量依赖性升高,在H1299-P53细胞中呈现与AKT相同的变化趋势,在H1299-175H细胞中4GY时表达量最高8GY最低。Caspase-3在H1299细胞中随着照射剂量的增加变化不明显,在H1299-P53细胞中呈现剂量依赖性升高,在H1299-175H细胞中,0 GY表达最高,4 GY表达最低,8 GY低于假照组但高于4 GY组。结论 X射线促进P53(-/-)的H1299细胞时发生自噬,而促进H1299-P53及H1299-175H中发生凋亡。
Objective To study the role of p53 gene in the ionizing radiation-induced death of non-small cell lung cancer cells. Methods H1299 (p53 - / -), H1299-P53 and H1299-175H cells were selected as research objects. Flow cytometry was used to detect the apoptosis rate. WESTERN BOLT method was used to detect the expression of related proteins. Results The apoptotic rates of H1299 (P53 - / -) cells after irradiation with 4GY and 8GY were 68.4% and 50% of that of the sham group, respectively. In H1299-P53 cells, the apoptosis rates of 4GY and 8GY irradiation were 1.7 times and 8.1 times that of the sham group respectively. In H1299-175H cells, the apoptotic rates of 4GY8 and GY after irradiation were 1.5 times and 2 times that of sham group respectively. The expression of LC3 protein showed a dose-dependent increase in H1299 and a dose-dependent reduction in H1299-175H. The highest expression of OGY, the lowest expression of 4 GY and the lower expression of 8 GY in H1299-P53 were observed . AKT expression decreased in a dose-dependent manner in H1299 cells. The expression of H1299-P53 increased in 4GY and decreased in 8GY, but still higher than that in sham group. AKT expression in H1299-175H cells decreased in a dose-dependent manner. The expression of MDM2 in H1299 increased in a dose-dependent manner, showing the same trend as that of AKT in H1299-P53 cells, and the highest expression level of 4GY in H1299-175H cells was the lowest. In H1299 cells, the expression of Caspase-3 did not change obviously with the increase of radiation dose, and showed a dose-dependent increase in H1299-P53 cells. The expression of Caspase-3 in H1299-175H cells was the highest, the lowest was 4 GY and the lowest was 8 GY In the sham group but higher than the 4 GY group. Conclusion X-ray can promote autophagy in P53 (- / -) H1299 cells and promote apoptosis in H1299-P53 and H1299-175H cells.