[1]尹　雷　王　鹏　侯崇显　李炎稳　周　东.靶向沉默Wip1基因表达对胶质瘤细胞增殖及放疗敏感性的作用[J].中国临床神经外科杂志,2018,(05):335-338.[doi:10.13798/j.issn.1009-153X.2018.05.010]
　YIN Lei,WANG Peng,HOU Chong-xian,et al.Effect of targeted silencing Wip1 gene on glioma cells proliferation and sensitivity to radiotherapy[J].,2018,(05):335-338.[doi:10.13798/j.issn.1009-153X.2018.05.010]
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靶向沉默Wip1基因表达对胶质瘤细胞增殖及放疗敏感性的作用()
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《中国临床神经外科杂志》[ISSN:1009-153X/CN:42-1603/TN]

卷:
期数:
2018年05期
页码:
335-338
栏目:
论著
出版日期:
2018-05-25

文章信息/Info

Title:
Effect of targeted silencing Wip1 gene on glioma cells proliferation and sensitivity to radiotherapy
文章编号:
1009-153X（2018）05-0335-04
作者:
尹　雷　王　鹏　侯崇显　李炎稳　周　东
作者单位：510515 广州，南方医科大学第二临床医学院（尹　雷、周东）；510080 广州，广东省人民医院、广东省医学科学院神经外科（尹雷、王　鹏、侯崇显、李炎稳、周　东）
Author(s):
YIN Lei1; 2;  WANG Peng2;  HOU Chong-xian2;  LI Yan-wen2;  ZHOU Dong1; 2
1. The Second School of Clinical Medicine, Southern Medical University, Guangzhou 510515, China; 2. Department of Neurosurgery, Guangdong General Hospital, Guangdong Academy of Medical Science, Guangzhou 510080, China
关键词:
胶质瘤细胞; 放疗; Wip1; p53; 基因沉默; 细胞增殖
Keywords:
Glioma;  U251 cells;  Wip1 gene;  Radiotherapy;  Silencing;  Proliferation;  Chk1;  Chk2;  Expression
分类号:
R 739.41; Q 812
DOI:
10.13798/j.issn.1009-153X.2018.05.010
文献标志码:
A
摘要:
目的 探讨靶向沉默Wip1基因表达对脑胶质瘤细胞增殖及放疗敏感性的影响。方法 体外培养人胶质瘤细胞株U251，用携带Wip1基因RNA干扰载体的慢病毒感染U251细胞沉默Wip1基因表达，然后对U251胶质瘤细胞进行放疗干预。根据对U251细胞处理方法分为4组：Wip1基因沉默后放疗组（IR+Wip1组）、单纯Wip1基因沉默组（Wip1组）、单纯放疗组（IR组）和NC组（空载体病毒感染U251细胞）。CCK-8法检测细胞的增殖，实时定量PCR检测细胞Chk1、Chk2 mRNA表达，免疫印迹检测细胞Chk1、Chk2蛋白表达。结果 U251细胞慢病毒感染后3~4 d，采用荧光显微镜观察，根据绿色荧光蛋白阳性表达判定感染效率，结果显示感染效率均90％以上。放疗后24、48、72、96、120 h，IR组和Wip1组增殖率均显著低于NC组（P<0.05），而IR+Wip1组细胞增殖率明显低于IR组和Wip1组（P<0.05）。放疗后24 h，IR+Wip1组Chk1 mRNA表达明显低于Wip1组（P<0.05），明显高于IR组和NC组（P<0.05）；IR+Wip1组Chk2 mRNA表达明显低于其他3组（P<0.05）。IR+Wip1组Chk1蛋白表达明显低于IR组和NC组（P<0.05），Chk2蛋白表达低于其他3组（P<0.05）。结论 靶向沉默 Wip1 基因表达可抑制胶质瘤细胞的增殖，并可能通过抑制 Chk1 、 Chk2 的蛋白表达增加胶质瘤细胞对放疗的敏感性。
Abstract:
Objective To investigate the effects of targeted silencing of Wip1 gene on glioma cells proliferation and sensitivity to radiotherapy. Methods Glioma U251 cells which were cultured were transfected with Wip1 RNAi lentiviral vector. Then the U251 cells were treated with ionizing radiation (IR) and divided into negative control (NC) group, IR group, targeted silencing Wip1 gene (Wip1) group and IR+Wip1 group. The proliferation activities of U251 cells, Chk1 and Chk2 mRNA expressions and Chk1 and Chk2 protein expressions were detected respectively by CCK-8 kit, real time PCR and Western-blot in all the groups. Results The proliferation rates in the Wip1 and the IR groups were significantly lower than that in NC group 24, 48, 72, 96 and 120 hours after the radiotherapy (P<0.05). The proliferation rate was significantly lower in the IR+Wip1 group than those in all the other groups 24, 72, 96 and 120 hours after the radiotherapy (P<0.05). The level of chk1 mRNA expression was significantly lower in IR+Wip1 group than that in Wip1 group and higher than those in IR and NC groups 24 hours after the radiotherapy (P<0.05). The level of chk2 mRNA expression was significantly lower in IR+Wip1 group than those in the other three groups (P<0.05).The level of Chk1 protein expression was significantly lower in IR+Wip1 group than those in IR and NC groups 24 hours after the radiotherapy (P<0.05). The level of Chk2 protein expression was significantly lower in the IR+Wip1 group than those in the other three groups (P<0.05). Conclusions It is suggested that targeted silencing Wip1 gene can inhibit the proliferation of glioma cells and may increase the sensitivity of glioma cells to radiotherapy by inhibiting the protein expressions of Chk1 and Chk2.

参考文献/References:

[1] Pechá?ková S, Burdová K, Macurek L. WIP1 phosphatase as pharmacological target in cancer therapy [J]. J Mol Med, 2017, 95(6): 589-599.
[2] Lowe J, Cha H, Lee MO, et al. Regulation of the Wip1 phos- phatase and its effects on the stress response [J]. Front Biosci (Landmark Ed), 2012, 17: 1480-1498.
[3] Goloudina AR, Kochetkova EY, Pospelova TV, et al. Wip1 phosphatase: between p53 and MAPK kinases pathways [J]. Oncotarget, 2016, 7(21): 31563-31571.
[4] Mirzayans R, Andrais B, Scott A, et al. Ionizing radiation- induced responses in human cells with differing TP53 status [J]. Int J Mol Sci, 2013, 14(11): 22409-22435.
[5] 王　鹏，饶　竟，杨海峰，等. 慢病毒介导的RNA干扰沉 默Wip1基因对人脑胶质母细胞瘤细胞生长的影响[J]. 中华实验外科杂志，2010，27（4）：481-483.
[6] Wang XW, Labussiere M, Valable S, et al. IDH1(R132H) mutation increases U87 glioma cell sensitivity to radiation therapy in hypoxia [J]. Biomed Res Int, 2014, 2014: 198697.
[7] Wang P, Rao J, Yang H, et al. PPM1D silencing by lentivi- ral-mediated RNA interference inhibits proliferation and invasion of human glioma cells [J]. J Huazhong Univ Sci Technolog Med Sci, 2011, 31(1): 94-99.
[8] Wang P, Rao J, Yang H, et al. Wip1 over-expression corre- lated with TP53/p14(ARF) pathway disruption in human astrocytomas [J]. J Surg Oncol, 2011, 104(6): 679-684.
[9] Rayter S, Elliott R, Travers J, et al. A chemical inhibitor of PPM1D that selectively kills cells overexpressing PPM1D [J]. Oncogene, 2008, 27(8): 1036-1044.
[10] Macurek L, Lindqvist A, Voets O, et al. Wip1 phosphatase is associated with chromatin and dephosphorylates gamma H2AX to promote checkpoint inhibition [J]. Oncogene, 2010, 29(15): 2281-2291.
[11] 凌　晖，文　玲，陈真伟，等. Chk1/2基因沉默对人胃癌 MGC803细胞增殖及周期的影响[J]. 中华肿瘤防治杂志， 2010，17（7）：481-484.
[12] Yoda A, Xu XZ, Onishi N, et al. Intrinsic kinase activity and SQ/TQ domain of Chk2 kinase as well as N-terminal domain of Wip1 phosphatase are required for regulation of Chk2 by Wip1 [J]. J Biol Chem, 2006, 281(34): 24847- 24862.
[13] Lu X, Nannenga B, Donehower LA. PPM1D dephosphory- lates Chk1 and p53 and abrogates cell cycle checkpoints [J]. Genes Dev, 2005, 19(10): 1162-1174.

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备注/Memo

备注/Memo:
基金项目：国家自然科学基金（81201995）；广东省医学科研基金（A2017183）；广东省自然科学基金（2015A030313532）；广东省科技计划项目（2014A020212678）
更新日期/Last Update: 2018-04-25