X线诱导肺腺癌SPC-A-1细胞发生Parthanatos

发布时间：2018-03-19 16:11

本文选题：AIF　切入点：X线辐射　出处：《第三军医大学学报》2015年16期 　论文类型：期刊论文

【摘要】：目的初步探讨X线诱导肺癌SPC-A-1细胞是否发生新的细胞死亡形式Parthanatos。方法以肺腺癌SPC-A-1细胞为研究对象,以1、2、4、6、8、10 Gy梯度剂量辐射细胞,平板克隆法测定X线辐射对人肺癌细胞SPC-A-1的半数致死剂量(LD50),WST-1法进行验证。LD50辐射细胞后4、8、12、24 h,RT-PCR法检测细胞AIF转录水平变化,Western blot检测细胞AIF蛋白表达变化,激光共聚焦观察细胞AIF向细胞核内转移情况,Western blot检测细胞辐射后不同时间AIF在线粒体蛋白组分和核蛋白组分中的分布变化。TUNEL法检测辐射后12 h细胞核DNA的断裂。结果 X线对SPC-A-1细胞的LD50大致为3 Gy。以3 Gy辐射细胞后不同时间AIF在转录、蛋白水平均无显著变化(P0.05)。辐射后12 h AIF由线粒体向细胞核转位,伴随细胞核的溶解、碎裂。亚细胞组分Western blot检测到12 h后AIF在线粒体蛋白组分中显著下降,由对照组的1.12±0.15下降至12 h组的0.21±0.07(P0.05),而在核蛋白组分中显著上升,由对照组的0.05±0.02上升至12 h组的0.45±0.07(P0.05),同时在固缩的细胞核中检测到DNA的断裂。结论 AIF核转位参与了X线诱导的细胞凋亡过程。辐射后12 h AIF由线粒体向细胞核转位显著增多,导致细胞核固缩、碎裂、溶解,细胞发生了Parthanatos。
[Abstract]:Objective to investigate whether a new cell death form Parthanatosin was found in lung cancer SPC-A-1 cells induced by X-ray. Methods SPC-A-1 cells from lung adenocarcinoma were studied and irradiated with a gradient dose of 810 Gy. The expression of AIF protein in human lung cancer cell line SPC-A-1 was detected by reverse transcription-polymerase chain reaction (RT-PCR) at 24 h after irradiation with LD50. The expression of AIF protein was detected by Western blot. Laser confocal scanning was used to observe the transfer of AIF into the nucleus. The distribution of AIF in mitochondria and nuclear protein at different time after irradiation was detected by Western blot. Tunel method was used to detect the break of nuclear DNA 12 h after irradiation. Results the LD50 of SPC-A-1 cells was about 3Gy. AIF was transcribed at different time after 3 Gy irradiation. There was no significant change in protein level (P 0.05). AIF translocated from mitochondria to nucleus at 12 h after irradiation, followed by nuclear dissolution and fragmentation. The subcellular component Western blot showed a significant decrease in AIF in mitochondrial protein component 12 h after irradiation. It decreased from 1.12 卤0.15 in control group to 0.21 卤0.07 P0.05 in 12h group, but increased significantly in nuclear protein component. From 0. 05 卤0. 02 in the control group to 0. 45 卤0. 07 P0. 05 in the 12 h group, DNA fragmentation was detected in the pyknotic nucleus. Conclusion the nuclear translocation of AIF is involved in the apoptosis induced by X ray. The translocation of AIF from mitochondria to the nucleus increases significantly at 12 h after irradiation. It causes the nucleus to shrink, break, and dissolve, and the cells develop Parthanatos.
【作者单位】：广东省人民医院(广东省医学科学院)肿瘤中心放疗科;广东医学院附属彭湃纪念医院肿瘤科;
【基金】：国家自然科学基金青年基金(81302033)~~
【分类号】：R734.2;R730.44

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