抑制蛋白磷酸酶2A对鼻咽癌放射敏感性的影响

doi:10.3969/j.issn.1004-616x.2016.04.003

癌变·畸变·突变 ›› 2016, Vol. 28 ›› Issue (4): 262-268.doi: 10.3969/j.issn.1004-616x.2016.04.003

抑制蛋白磷酸酶2A对鼻咽癌放射敏感性的影响

吕鹏^1,2,6, 满其忠³, 王越⁴, 庄正平⁵, 王爱平¹, 曾益新⁶

1. 中国医学科学院北京协和医学院新药安全评价研究中心, 北京 100050;
2. 中华医学会杂志社, 中华医学杂志英文版, 北京 100710;
3. 山东万杰医学院解剖教研室, 山东淄博 255213;
4. 国家食品药品监督管理总局, 中国食品药品检定研究院, 医疗器械标准管理研究所, 北京 100050;
5. 美国国立卫生研究院神经失调与中风研究所神经外科研究室, 美国贝塞斯达 20892;
6. 中国医学科学院北京协和医学院肿瘤医院肿瘤研究所, 北京 100021

收稿日期:2016-04-16 修回日期:2016-06-04 出版日期:2016-07-31 发布日期:2016-07-31
通讯作者: 王爱平,E-mail:wangaiping@imm.ac.cn;曾益新,E-mail:zengyix@mail.sysu.edu.cn E-mail:zengyix@mail.sysu.edu.cn
作者简介:吕鹏,E-mail:lvpeng51@foxmail.com;王越,E-mail:wang_yue_nifdc@sina.com
基金资助:
国家自然科学基金项目（81502389）

Effects of protein phosphatase 2A inhibition on radiosensitivity of nasopharyngeal carcinoma

LÜ Peng^1,2,6, MAN Qizhong³, WANG Yue⁴, ZHUANG Zhengping⁵, WANG Aiping¹, ZENG Yixin⁶

1. The New Drug Safety Evaluation Center, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China;
2. Chinese Medical Journal, Chinese Medical Association Publishing House, Beijing 100710, China;
3. Department of Anatomy, Shandong Wanjie Medical College, Zibo 255213, Shandong, China;
4. Institute for Medical Device Standardization Administration, National Institutes for Food and Drug Control, China Food and Drug Administration, Beijing 100050, China;
5. Surgical Neurology Branch, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda 20892, USA;
6. Cancer Institute and Hospital, Chinese Academy of Medical Sciences, Beijing 100021, China

Received:2016-04-16 Revised:2016-06-04 Online:2016-07-31 Published:2016-07-31

摘要/Abstract

摘要： 目的：研究放射处理条件下蛋白磷酸酶2A （PP2A）的抑制对鼻咽癌CNE2细胞及其裸鼠移植瘤放射增敏的效果。方法：建立鼻咽癌细胞CNE2体外实验与裸鼠体内移植瘤模型，随机分为空白对照组（PBS）、单纯去甲斑蝥素药物组（NCTD，体外40μmol/L，体内27μmol/kg）、单纯放射组（体外8 Gy，体内20 Gy）及联合处理组。采用四甲基噻唑蓝（MTT）、免疫共沉淀、流式细胞术等方法研究去甲斑蝥素和或放射处理对鼻咽癌CNE2细胞PP2A活性、细胞周期、细胞凋亡及裸鼠体内移植瘤的影响。结果：体外和体内实验均显示，单纯NCTD处理组5 h后PP2A活性均被抑制，约为对照组的70%，而单纯放射组处理5 h后PP2A蛋白活性明显升高，分别约为对照组的210%（体外）和165%（体内），差异均具有统计学意义（P<0.05）。单纯NCTD处理组、单纯放射组均可不同程度的引起CNE2细胞G2/M期阻滞及细胞凋亡率的增加，与对照组间的差异均具有统计学意义（P<0.05）。联合处理组与对照组比较，PP2A的活性明显降低，分别约为对照组的80%（体外细胞实验）和72%（体内裸鼠实验）；同时G2/M期细胞显著增多（87.88%±2.10%），细胞凋亡率也明显提高（77.15%±7.62%）；裸鼠移植瘤抑瘤率达到87.98%，其差异均具有统计学意义（P均<0.05）。移植瘤组织病理学观察发现联合处理组大量细胞坏死，部分融合成片，核膜核仁破碎，在肿瘤组织细胞非死亡区可见到被瘤细胞吞噬形成的凋亡小体。结论：PP2A很有可能成为提高鼻咽癌临床放射治疗作用的增敏新靶点。

关键词: 蛋白磷酸酶2A, 去甲斑蝥素, 放射增敏, 鼻咽癌

Abstract: OBJECTIVE: To investigate the radiosensitizing effect of protein phosphatase 2A(PP2A) inhibition on nasopharyngeal carcinoma cell line CNE2 and its xenografts. METHODS: The cells and subcutaneous xenografts in BALB/c nude mice were randomly divided into control, norcantharidin (40 μmmol/L in vitro, 27 μmol/kg in vitro), irradiation (8 Gy in vitro, 20 Gy in vitro), and combination of norcantharidin and irradiation groups. MTT, Co-immunoprecipitation and flow cytometry were used to examine the effect of PP2A inhibition, cell cycle analysis, apoptosis of CNE2 and its xenografts by norcantharidin and (or) irradiation. RESULTS: We measured PP2A activity in CNE2 cells and xenografts 5 hours after exposure to 8 Gy radiation in vitro and 20 Gy radiation in vivo, with and without prior exposure to NCTD. Compared with control group, NCTD alone was associated with decreasedin PP2A activity of 70% in vitro and in vivo both. Radiation alone was associated with increased in PP2A activity of 210% and 165% in vitro and in vivo, respectively. The cell cycle arrested in G2/M and increasing of apoptotic ratio in CNE2 cells were significantly different between the control and the both two treatment alone groups (P<0.05). Compared with control group, the combination of 8 Gy and 40 μmol/L norcantharidin produced significant inhibition of PP2A(80% in vitro and 72% in vitro compared to control), accumulation of cells in G2/M-phase (87.88%±2.10%) and more apoptosis (77.15%±7.62%) in CNE2 cell lines compared to norcantharidin alone and radiation alone. Norcantharidin in combination with radiation produced the greatest effects on tumor growth slowing and decreasing tumor weight by 87.98% relative to control in CNE2 xenografts(compared to control, P<0.05). Furthermore, more apoptosis, necrotic cells and formation of apoptotic bodies were found in pathological section in norcantharidin plus radiotherapy group. CONCLUSION: PP2A might be a potential target for sensitization of nasopharyngeal carcinoma to radiatotherapy.

Key words: protein phosphatase 2A, norcantharidin, radiosensitizion, nasopharyngeal carcinoma

中图分类号:

R739.6

吕鹏, 满其忠, 王越, 庄正平, 王爱平, 曾益新. 抑制蛋白磷酸酶2A对鼻咽癌放射敏感性的影响[J]. 癌变·畸变·突变, 2016, 28(4): 262-268.

LÜ Peng, MAN Qizhong, WANG Yue, ZHUANG Zhengping, WANG Aiping, ZENG Yixin. Effects of protein phosphatase 2A inhibition on radiosensitivity of nasopharyngeal carcinoma[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2016, 28(4): 262-268.

参考文献

[1] Hsu WL,Chen JY,Chien YC,et al. Independent effect of EBV and cigarette smoking on nasopharyngeal carcincma:a 20- year follow-up study on 9622 males without family history in Taiwan cancer epidemiol[J]. Biomarkers Prev,2009,18(4):1218-1226.
[2] 王中卫,王亚利,金迎迎,等. 蛋白组学分析放射抗拒性鼻咽癌细胞差异表达蛋白[J]. 西安交通大学学报:医学版,2014,35(6):810-815.
[3] 张建庆,陈惠,杨杰,等. 周剂量奈达铂联合调强放疗对复发鼻咽癌的近期疗效分析[J].现代生物医学进展,2013,13(20):3881-3884.
[4] Zhang L,Chen QY,Liu H,et al.Emerging treatment options for nasopharyngeal carcinoma[J]. Drug Des Devel Ther,2013,7(1):37-52.
[5] Tham IW,Hee SW,Yap SP. Retropharyngeal nodal metastasis related to higher rate of distant metastasis in patient with N0 and N1 nasopharyngeal cancer[J]. Head Neck,2009,31(4):468-474.
[6] Cheng G,Wei L,Xiurong W,et al. IL-17 stimulates migration of carotid artery vascular smooth muscle cells in an MMP-9 dependent manner via p38MAPK and ERK1/2-dependent NF-kappaB and AP-1 activation[J]. Cell Mol Neurobiol,2009,29(8):1161-1168.
[7] Woo MM,Salamanca CM,Symowicz J,et al.SV40 early genes induce neoplastic properties in serous berdedine ovarian tumor cells[J]. Gynecol Oncol,2008,111(1):125-131.
[8] 李天祝,向本琼. 蛋白磷酸酶PP2A的结构及其肿瘤抑制因子功能[J]. 生物化学与生物物理进展,2009,36(2):133-142.
[9] Zhuang Z,Lu J,Lonser R,et al. Enhancement of cancer chemotherapy by simultaneously altering cell cycle progression and DNA-damage defenses through global modification of the serine/threonine phospho-proteome[J]. Cell Cycle,2009,8(20):3303-3306.
[10] 毛燕萍,李文斐,陈磊,等. 鼻咽癌2008分期的临床验证[J]. 癌症,2009,28(10):1022-1028.
[11] Mumby M. PP2A:unveiling a reluctant tumor suppressor[J]. Cell,2007,130(1):21-24.
[12] Eichhorn PJ,Creyghton MP,Bernards R. Protein phosphatase 2A regulatory subunits and cancer[J]. Biochim Biophys Acta,2009,1795(1):1-15.
[13] Farkas I,Dombrádi V,Miskei M,et al. Arabidopsis PPP family of serine/threonine phosphatases[J]. Trends Plant Sci, 2007,12(4):169-176.
[14] Hofstetter CP,Burkhardt JK,Shin BJ,et al. Protein phosphatase 2A mediates dormancy of glioblastoma multiforme-derived tumor stem-like cells during hypoxia[J]. PloS ONE,2012,7(1):30059.
[15] Wei D,Parsels L,Karnak D,et al. Inhibition of protein phosphatase 2A radiosensitizes pancreatic cancers by modulating CDC25C/CDK1 and homologous recombination repair[J]. Clin Cancer Res,2013,19(16):4422-4432.
[16] Yan Y,Cao PT,Greer PM,et al. Protein phosphatase 2A hasan essential role in the activation of gamma-irradiation induced G2/M checkpoint response[J]. Oncogene,2010,29(30):4317-4329.
[17] Kovach JS,Johnson F. Oxabicycloheptanes and oxabicylco-heptenes,their preparation and use:us,7998957[P/OL]. 2011-08-16[2015-06-15]. http://patent.ipexl.com/US/07998957.html.
[18] Lu J,Kovach JS,Johnson F,et al. Inhibition of serine/threonine phosphatase PP2A enhances cancer chemotherapy by blocking DNA damage induced defense mechanisms[J]. PNAS,2009,106:11697-11702.
[19] Wei D,Parsels LA,Karnak D,et al. Inhibition of protein phosphatase 2A radiosensitizes pancreatic cancers by modulating CDC25C/CDK1 and homologous recombination repair[J]. Clin Cancer Res,2013,19(16):4422-4432.
[20] Martiniova L,Lu J,Chiang J,et al. Pharmacologic modulation of serine/threonine phosphorylation highly sensitizes PHEO in a MPC cell and mouse model to conventional chemotherapy[J]. PloS ONE,2011,6(2):14678.
[21] Li W,Chen Z,Gong FR,et al. Growth of the pancreatic cancer cell line PANC-1 is inhibited by protein phosphatase 2A inhibitors through over activation of the c-Jun N-terminal kinase pathway[J]. European J Cancer,2011,47(17):2654-2664.
[22] Deng L,Dong J,Wang W. Exploiting protein phosphatase inhibitors based on cantharidin analogues for cancer drug discovery[J]. Mini-Rev Med Chem,2013,13(8):1166-1176.
[23] Lankoff A,Bialczyk J,Dziga D,et al. The repair of gamma-radiation-induced DNA damage is inhibited by microcystin-LR,the PP1 and PP2A phosphatase inhibitor[J]. Mutagenesis,2006,21(1):83-90.

抑制蛋白磷酸酶2A对鼻咽癌放射敏感性的影响

Effects of protein phosphatase 2A inhibition on radiosensitivity of nasopharyngeal carcinoma

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