γ射线对小鼠胸腺淋巴细胞微丝形态及丝切蛋白磷酸化的影响

doi:10.3969/j.issn.1004-616x.2018.04.001

癌变·畸变·突变 ›› 2018, Vol. 30 ›› Issue (4): 249-253,257.doi: 10.3969/j.issn.1004-616x.2018.04.001

• 论著 • 下一篇

γ射线对小鼠胸腺淋巴细胞微丝形态及丝切蛋白磷酸化的影响

齐雪松, 王春燕, 李辰, 佟鹏, 邵帅, 曲功霖, 苟巧, 苏旭

中国疾病预防控制中心辐射防护与核安全医学所, 辐射防护与核应急中国疾病预防控制中心重点实验室, 北京 100088

收稿日期:2018-02-22 修回日期:2018-03-09 出版日期:2018-07-30 发布日期:2018-07-30
通讯作者: 苏旭,E-mail:suxu@nirp.chinacdc.cn E-mail:suxu@nirp.chinacdc.cn
作者简介:齐雪松,E-mail:cedar121@sina.com
基金资助:
国家自然科学基金青年基金（81402632）

Effect of γ-ray on microfilament morphology and cofilin phosphorylation of thymus lymphocytes of mice

QI Xuesong, WANG Chunyan, LI Chen, TONG Peng, SHAO Shuai, QU Gonglin, GOU Qiao, SU Xu

Key Laboratory of Radiological Protection and Nuclear Emergency, National Institute for Radiological Protection, Chinese Center for Disease Control and Prevention, Beijing 100088, China

Received:2018-02-22 Revised:2018-03-09 Online:2018-07-30 Published:2018-07-30

摘要/Abstract

摘要： 目的：通过观察γ射线诱导小鼠胸腺淋巴细胞的微丝形态变化和对丝切蛋白（cofilin）磷酸化的影响，探讨cofilin及相关蛋白在辐射损伤中的作用。方法：将36只BALB/c小鼠按对照组（0 h），照射后3、6、9、12和24 h时间点随机分为6组，每组6只，除对照组外，其余各组均给予2 Gy γ射线诱导，根据cofilin蛋白表达规律，确定取材时间。另将24只BALB/c小鼠按γ射线诱导剂量随机分为未照射（0 Gy）组、2、6和10 Gy组共4组，照射3 h后取材，观察淋巴细胞微丝形态，进行cofilin蛋白1（cofilin 1）、磷酸化cofilin蛋白（p-cofilin）、LIM激酶1（LIMK1）、TES激酶2（TESK2）和Slingshot家族的磷酸酶2（SSH2）表达检测。结果：BALB/c小鼠在2 Gy γ射线照射3 h后胸腺淋巴细胞cofilin基因的mRNA和蛋白表达均显著下调（P < 0.05），因此后续实验选择照后3 h为取材时间。通过对cofilin及相关蛋白表达的检测，发现随着γ射线照射剂量的增加，胸腺淋巴细胞cofilin的表达逐渐增加，与对照组相比，以10 Gy组增加最为明显（P < 0.05）；而p-cofilin的表达则减少，10 Gy组减少最为明显（P < 0.05）。LIMK1、TESK2和SSH2的表达在照射后无显著变化（P > 0.05）。结论：随着照射剂量的增加，更多的SSH2使p-cofilin的Ser3位点去磷酸化，胞质内cofilin的量增多，参与到微丝骨架组装的过程中，细胞的迁移能力增强。

关键词: γ射线, 胸腺淋巴细胞, 微丝骨架, 丝切蛋白

Abstract: OBJECTIVE: To explore the role of cofilin for radiation damage via changes of microfilament morphology and cofilin phosphorylation which were induced by γ-ray in thymus lymphocytes of mice. METHODS: 36 mice were grouped randomly according to the time after radiation into control (0 h),3,6,9,12 and 24 h. Thymus lymphocytes were collected at the mentioned time points after exposure to 2 Gy γ-rays. Cofilin 1 in lymphocytes was detected by Western blot to make decision for the suitable time point. 24 mice were grouped randomly according to the dose of γ-ray,including 4 groups of control without radiation (0 Gy),2,6,10 Gy. Three hours after γ-ray exposure,lymphocytes were collected to observe some changes of microfilament morphology and cofilin 1 phosphorylation,including the expression of cofilin 1,p-cofilin,LIMK1,TESK2 and SSH2. RESULTS: mRNA and protein expression of cofilin were down regulated (P < 0.05),the time point of 3 h after γ-ray exposure was the suitable time for collecting thymus lymphocytes to test the expression of cofilin and its related proteins. The results show that expression of cofilin increased with increasing doses of radiation,especially the group of 10 Gy was significantly different (P < 0.05). On the contrary,expression of p-cofilin was reduced,especially the group of 10 Gy was significantly different (P < 0.05). The expression of LIMK1,TESK2 and SSH2 looked similar to controls(P > 0.05). CONCLUSION: The increased expression of SSH2 caused p-cofilin dephosphorylating at Ser3,indicating the quantity of cofilin 1 increased in cytoplasm to take part in microfilament assembly. As a result,movement ability of the lymphocytes was enhanced.

Key words: γ-ray, thymus lymphocytes, microfilament, cofilin

中图分类号:

R994.6

齐雪松, 王春燕, 李辰, 佟鹏, 邵帅, 曲功霖, 苟巧, 苏旭. γ射线对小鼠胸腺淋巴细胞微丝形态及丝切蛋白磷酸化的影响[J]. 癌变·畸变·突变, 2018, 30(4): 249-253,257.

QI Xuesong, WANG Chunyan, LI Chen, TONG Peng, SHAO Shuai, QU Gonglin, GOU Qiao, SU Xu. Effect of γ-ray on microfilament morphology and cofilin phosphorylation of thymus lymphocytes of mice[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2018, 30(4): 249-253,257.

参考文献

[1] 齐雪松,苏旭. 微丝骨架在T细胞活化中的作用[J]. 癌变·畸变·突变,2012,24(3):238-240.
[2] 包志瑶,周新,韩黎. 丝切蛋白与肺部疾病的研究进展[J]. 国际呼吸杂志,2014,34(17):1353-1356.
[3] BERNATEIN B W,BANBURG J R. ADF/Cofilin:a function node in cell biology[J]. Trends Cell Biol,2010,20(4):187-195.
[4] OSTROWSKA Z,MORACZEWSKA J. Cofilin-a protein controlling dynamics of actin filaments[J]. Postepy Hig Med Dosw(online),2017,71(10):339-351.
[5] KANELLOS G,FRAME M C. Cellular functions of the ADF/cofilin family at a glance[J]. J Cell Sci,2016,129(17):3211-3218.
[6] POLLARD T D. What we know and do not know about actin[J]. Handb Exp Pharmacol,2017,235:331-347.
[7] HAIBO W,LIDE T,FENG J,et al. Cofilin 1 induces the epithelial-mesenchymal transition of gastric cancer cells by promoting cytoskeletal rearrangement[J]. Oncotarget,2017,8(24):39131-39142.
[8] SU J,ZHOU Y,PAN Z,et al. Downregulation of LIMK1-ADF/cofilin by DADS inhibits the migration and invasion of colon cancer[J]. Sci Rep,2017,7:45624.
[9] 齐雪松,王春燕,郝述霞,等. γ射线诱导小鼠胸腺淋巴细胞中Arp2、Arp3和cofilin基因表达变化的研究[J]. 癌变·畸变·突变,2011,23(6):421-424.
[10] SAN M A,LEE M Y,WILLIAMS H C,et al. Dual regulation of cofilin activity by LIM kinase and Slingshot-1L phosphatase controls platelet-derived growth factor-induced migration of human aortic smooth muscle cells[J]. Circ Res,2008,102(4):432-438.
[11] PRUNIER C,PRUDENT R,KAPUR R,et al. LIM kinases:cofilin and beyond[J]. Oncotarget,2017,8(25):41749-41763.
[12] AGGELOU H,CHADLA P,NIKOU S,et al. LIMK/cofilin pathway and Slingshot are implicated in human colorectal cancer progression and chemoresistance[J]. Virchows Arch,2018. doi:10.1007/s00428-018-2298-0.
[13] SIMHADRI P K,MALWADE R,VANKA R,et al. Dysregulation of LIMK-1/cofilin-1 pathway:A possible basis for alteration of neuronal morphology in experimental cerebral malaria[J]. Ann Neurol,2017,82(3):429-443.
[14] ZHOU T,WANG CH,YAN H,et al. Inhibition of the Rac1-WAVE2-Arp2/3 signaling pathway promotes radiosensitivity via downregulation of cofilin-1 in U251 human glioma cells[J]. Sci Rep,2016,13(5):4414-4420.
[15] SHI C,CAI Y,LI Y,et al. Yap promotes hepatocellular carcionoma metastasisi and mobilinzation via governing coilin/F-actin/lamellipodium axis by regulation of JNK/Binp3/SERCA/CaMKⅡ pathways[J]. Redox Biol,2018,14(1):59-71.
[16] OHASHI K,SAMPEI K,NAKAGAWA M,et al. Damnacanthal,an effective inhibitor of LIM-kinase,inhibits cell migration and invasion[J]. Mol Biol Cell,2014,25(6):828-840.
[17] NAOUN G E,MORKOS M,KIM B,et al. Novel targeted therapies and immunotherapy for advanced thyroid cancers[J]. Mol Cancer,2018,17(1):51.

γ射线对小鼠胸腺淋巴细胞微丝形态及丝切蛋白磷酸化的影响

Effect of γ-ray on microfilament morphology and cofilin phosphorylation of thymus lymphocytes of mice

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 10

编辑推荐

Metrics

本文评价

[1]	王春燕, 佟鹏, 李辰, 邵帅, 曲功霖, 苟巧, 王成芳, 齐雪松. 纳米氧化铈颗粒对人肺腺癌耐顺铂细胞辐射敏感性的影响[J]. 癌变·畸变·突变, 2019, 31(6): 464-468.
[2]	孙鸽, 秦秀军, 王永丽, 李建国, 周朝东, 安全, 闻建华. ⁶⁰Co γ射线对雄性比格犬睾酮分泌及睾丸组织病理学的影响[J]. 癌变·畸变·突变, 2016, 28(6): 443-445,452.
[3]	张伟，秦秀军，许超琪，李炜宾，尹晶晶，袁慧，李建国. γ射线照后不同时间点人肝细胞基因差异表达谱的研究[J]. , 2011, 23(6): 416-420.
[4]	王卉，封江彬，陆雪，张德钦，张庆召，高玲，陈德清，刘青杰. 60Co γ射线诱导人淋巴细胞线粒体COX II 基因表达改变的研究[J]. , 2011, 23(6): 437-441.
[5]	齐雪松，王春燕，郝述霞，苟巧，佟鹏，苏旭. γ射线诱导小鼠胸腺淋巴细胞中 Arp2、 Arp3和cofilin 基因表达的变化[J]. , 2011, 23(6): 421-424.
[6]	薛振伟，周小林，崔梅萍. γ射线辐照对BEP2D细胞蛋白质组的影响[J]. , 2011, 23(6): 425-428.
[7]	王　卉,李玉文,封江彬,陆　雪,范　莉,陈德清,赵　骅,刘青杰. 60Co γ射线诱导人淋巴细胞GDF15mRNA表达水平的变化[J]. , 2010, 22(5): 352-355.
[8]	吴丛梅;黄天华;谢庆东;李德锐;陈秀如;吴德生;徐小虎. 重组质粒pEgr-P１6在Ec9706细胞中的辐射诱导凋亡作用[J]. , 2004, 16(3): 129-131.
[9]	刘建香;苏　旭. 电离辐射调控脂质体介导的TNFα基因在HeLa细胞中的表达[J]. , 2004, 16(2): 89-91.
[10]	吴丛梅1,黄天华1，吴德生,谢庆东1 ,徐小虎2. pEgr-P16重组质粒的构建及其在EC9706细胞中的辐射诱导表达[J]. , 2003, 15(4): 203-204.