盐诱导激酶2对电离辐射诱导的自噬与凋亡的影响

辐射防护 ›› 2017, Vol. 37 ›› Issue (3): 214-222.

盐诱导激酶2对电离辐射诱导的自噬与凋亡的影响

李超¹, 李忠秋^2,3, 李雪萍^1,3, 杨洋^2,3, 曾妍², 潘秀颉³, 杨陟华³, 朱茂祥³, 顾永清³

1.石河子大学生命科学学院, 新疆石河子 832003;
2.石河子大学医学院, 新疆石河子 832003;
3.军事医学科学院放射与辐射医学研究所, 北京 100850

收稿日期:2016-12-09 出版日期:2017-05-20 发布日期:2021-11-11
通讯作者: 顾永清。E-mail:yqgu96@163.com
作者简介:李超(1990—),男,2014年6月毕业于延安大学生物科学专业,现为石河子大学生物化学与分子生物学专业硕士研究生。E-mail:13681321363@163.com
基金资助:
国家自然科学基金资助项目(No.31470827; No.81472910; No.81560473; No.31270894)。

Effects of salt-inducible kinase 2 on autophagy and apoptosis induced by ionizing radiation

Li Chao¹, Li Zhongqiu^2,3, Li Xueping^1,3, Yang Yang^2,3, Zeng Yan², Pan Xiujie³, Yang Zhihua³, Zhu Maoxiang³, Gu Yongqing^3*

1. College of Life Sciences, Shihezi University, Xinjiang Shihezi 832003;
2. College of Medicine, Shihezi University, Xinjiang Shihezi 832003;
3. Department of Radiation Toxicology and Oncology, Beijing Institute of Radiation Medicine, Beijing 100850

Received:2016-12-09 Online:2017-05-20 Published:2021-11-11

摘要/Abstract

摘要： 为探索盐诱导激酶2(salt-inducible kinase 2, SIK2)对电离辐射引起的自噬与凋亡的影响,以期为肿瘤的放射治疗提供新的思路,以慢病毒颗粒LV-GFP和LV-SIK2体外转染HeLa细胞,48 h后用荧光显微镜观察转染是否成功,RT-PCR和Western blot检测转染细胞SIK2的干扰效果。通过Western blot检测SIK2低表达对8 Gy ⁶⁰Co γ射线照射下细胞中自噬相关蛋白LC3-Ⅱ 和P62表达的影响来观察自噬的变化;利用流式细胞术来检测SIK2低表达对8 Gy ⁶⁰Co γ射线照射下细胞凋亡的影响。结果表明,SIK2低表达的稳转细胞系被成功构建;8 Gy γ射线照射后,SIK2低表达细胞系LC3-Ⅱ表达下调,P62表达上调,表明自噬被减弱;流式细胞术检测发现SIK2低表达细胞系在照后12 h FITC⁺PI^-细胞比例就显著增加,表明SIK2低表达增加了细胞凋亡。因此,SIK2低表达会导致电离辐射引起的细胞自噬大大减弱,凋亡显著增加。

关键词: 盐诱导激酶2(SIK2), 电离辐射, 自噬, 凋亡, 肿瘤放疗

Abstract: The purpose of this study is to explore the effects of salt-inducible kinase 2 (SIK2) on autophagy and apoptosis induced by ionizing radiation in the hope of providing a new idea for tumor radiotherapy. Lentivirus LV-GFP and LV-SIK2 were transfected into HeLa cells in vitro. The success of transfection was examined by fluorescence microscopy after 48 h. Real-time PCR and Western blot were used to measure the efficiency of the SIK2 knockdown. After exposure to 8 Gy ⁶⁰Co γ irradiation, the effects of the SIK2 knockdown on autophagy related protein LC3-II and on the P62 expression were examined by Western blot to observe the changes of autophagy. Flow cytometry was performed to examine the effect of the SIK2 knockdown on the apoptosis after 8 Gy ⁶⁰Co γ irradiation. The results showed that the SIK2 knockdown of stably transfected cell line was established successfully. In the down-regulating SIK2 cell line after 8 Gy γ irradiation, the expression of LC3-II was down-regulated while the P62 expression was up-regulated, indicating that autophagy was reduced. Flow cytometric analysis showed that FITC⁺/PI^- cells ratio was already increased significantly at 12 h post irradiation in the down-regulating SIK2 cell line, indicating that the SIK2 knockdown increased the cell apoptosis. In conclusion, the SIK2 knockdown could lead to a significant increase of cell apoptosis induced by ionizing radiation.

Key words: SIK2, ionizing radiation, autophagy, apoptosis, tumor radiotherapy

中图分类号:

Q691

李超, 李忠秋, 李雪萍, 杨洋, 曾妍, 潘秀颉, 杨陟华, 朱茂祥, 顾永清. 盐诱导激酶2对电离辐射诱导的自噬与凋亡的影响[J]. 辐射防护, 2017, 37(3): 214-222.

Li Chao, Li Zhongqiu, Li Xueping, Yang Yang, Zeng Yan, Pan Xiujie, Yang Zhihua, Zhu Maoxiang, Gu Yongqing. Effects of salt-inducible kinase 2 on autophagy and apoptosis induced by ionizing radiation[J]. RADIATION PROTECTION, 2017, 37(3): 214-222.

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