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《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]

卷:

21卷

期数:

2015年06期

页码:

461-465

栏目:

基础医学

出版日期:

2015-11-20

文章信息/Info

Title:

DZNEP inhibits cell proliferation and induce s apoptosis by targeting EZH2 in human head and neck squamous cell carcinoma in vitro and vivo

文章编号:

1006-8147(2015)06-0465-05

作者:

孔令平 ; 周 旋;  孙姗姗 ; 黄媛媛;  张 仑

(天津医科大学肿瘤医院颌面部耳鼻喉科,国家肿瘤临床医学研究中心,天津市“肿瘤防治”重点实验室 ,天津 300060)

Author(s):

KONG Ling-ping;  ZHOU Xuan;  SUN Shan-shan;  HUANG Yuan-yuan;  ZHANG Lun

(Department of Maxillofacial & Ear, Nose, and Throat Oncology, Cancer Institute and Hospital , Tianjin Medical University ,National Clinical Research Center of Cancer. Key Laboratory of Cancer Prevention and Therapy, Tianjin. Tianjin 300060, China)

关键词:

头颈部鳞状细胞癌' target="_blank" rel="external">">头颈部鳞状细胞癌; EZH2; 细胞凋亡; 细胞增殖

Keywords:

head and neck squamous cell carcinoma;  EZH2;  apoptosis;  proliferation

分类号:

R737.9

DOI:

-

文献标志码:

A

摘要:

目的： 探讨Zeste同源序列2的增强子(EZH2)的小分子抑制剂DZNEP抑制人头颈部鳞状细胞癌细胞Tb3.1增殖、诱导凋亡的效果与机制。方法： 甲基噻唑基四唑（MTT）法测细胞对DZNEP的半数致死量（IC50）、细胞的增殖能力；流式细胞术检测细胞凋亡；平板克隆法检测细胞形成克隆能力；JC-1荧光探针染色法检测细胞线粒体膜电位；蛋白质印迹法(WB)检测EZH2、细胞增殖核抗原(Ki-67)、B细胞淋巴瘤2 (Bcl-2、BAX)、活化型含半胱氨酸的天冬氨酸蛋白水解酶3 (CCASP-3）的表达。体内实验裸鼠皮下荷瘤模型，通过免疫组织化学染色及原位末端标记（TUNEL）法检测DZNEP对细胞增殖、凋亡的作用。结果： 经DZNEP处理后，EZH2蛋白表达降低; MTT法检测DZNEP明显抑制鳞癌细胞Tb3.1的增殖，且具有浓度和时间依赖性；流式细胞术显示DZNEP可诱导细胞凋亡；平板克隆实验表明DZNEP能够明显抑制克隆形成能力，抑制细胞增殖；JC-1荧光探针染色法显示DZNEP可以改变细胞的线粒体膜电位；WB 显示，DZNEP可增加促凋亡基因（CCASP-3、BAX）的表达，抑制Ki-67、Bcl-2的表达水平。体内实验观察结束时，DZNEP处理组肿瘤体积较对照组明显减小。免疫组织化学染色结果示，DZNEP组中BAX、CCASP-3表达增加，Ki-67、Bcl-2表达减少；TUNEL结果显示，DZNEP组比DMSO 组肿瘤细胞凋亡指数上升。结论： DZNEP通过抑制EZH2表达在体外抑制Tb3.1细胞增殖并诱导凋亡；为进一步探讨EZH2参与人头颈部鳞状细胞癌发生发展的分子机制提供科学实验依据。

Abstract:

Objective ： To investigate the anti-tumour molecular mechanism of DZNEP in human head and neck squamous cell carcinoma. Methods： Tb3.1 human head and neck squamous cell carcinoma cell lines were employed. DZNEP was used to suppress the expression of EZH2. Methyl thiazolyl tatrozolium (MTT) assay was applied to determine IC50 and cell survival. Flow cytometry (FCM) was used to measure cell apoptosis. JC-1 fluorescence was employed to determine MMP. The expression level of EZH2 , antigen 67 (Ki-67), B cell lymphoma 2 (Bcl-2、BAX) and cleaved cysteinyl aspartate specific proteinase-3 (CCASP-3) were examined by Western blotting. Results：Compared with control and normal control cells, DZNEP inhibited EZH2 expression and affected cell proliferation. Apoptosis rate was elevated by DZNEP. Clone formation assay suggested that cell clone numbers were significantly reduced by DZNEP. BAX and CCASP-3 expression were enhanced by DZNEP treatment, while Ki-67 and Bcl-2 expression were suppressed. In vivo, the tumor volume after treatment with DZNEP was significantly smaller than Control groups. Immunological Histological Chemistry suggested that BAX and CCASP-3 protein expression was up-regulated while Ki-67 and Bcl-2 protein of tumor tissue was down-regulated after treated DZNEP as compared to control groups. TUNEL assay indicated that apoptosis index was increased after treatment with DZNEP compared with control groups. Conclusion：DZNEP modulates proliferation and apoptosis of Tb3.1 cancer cell by inhibiting EZH2, which might promote further research in head and neck squamous cell carcinoma treatment.

参考文献/References:

[1].1]Ezhkova E, Pasolli H A, Parker J S, et al. Ezh2 orchestrates gene expression for the stepwise differentiation of Tissue-Specific stem cells[J]. Cell, 2009, 136(6): 1122
[2].[2]Vire E, Brenner C, Deplus R, et al. The polycomb group protein EZH2 directly controls DNA methylation[J]. Nature, 2006, 439(7078): 871
[3].[3]Strojan P, Vermorken J B, Beitler J J, et al. Cumulative cisplatin dose in concurrent chemoradiotherapy for head and neck Cancer:A systematic review[J]. Head Neck, 2015，?[Epub ahead of print]
[4]. [4]Safdari Y, Khalili M, Farajnia S A, et al. Recent advances in head and neck squamous cell carcinoma - A review[J]. Clin Biochem, 2014, 47(13/14): 1195
[5].[5]Burz C, Berindan-Neagoe I, Balacescu O, et al. Apoptosis in Cancer:key molecular signaling pathways and therapy targets[J]. Acta Oncol, 2009, 48(6): 811
[6].[6]Nakagawa S, Sakamoto Y, Okabe H, et al. Epigenetic therapy with the histone methyltransferase EZH2 inhibitor 3-deazaneplanocin A inhibits the growth of cholangiocarcinoma cells[J]. Oncol Rep, 2014, 31(2): 983
[7].[7]Wang Cheng, Liu Xi QIANG, Chen Zu JIAN, et al. Polycomb group protein EZH2-mediated E-cadherin repression promotes metastasis of oral tongue squamous cell carcinoma[J]. Mol Carcinog, 2013, 52(3): 229
[8].[8]Cao Wei, Feng Z, Cui Zhi BIN, et al. Up-regulation of enhancer of zeste homolog 2 is associated positively with cyclin D1 overexpression and poor clinical outcome in head and neck squamous cell carcinoma[J]. Cancer, 2012, 118(11): 2858
[9].[9]Fiskus W, Rao R, Balusu R, et al. Superior efficacy of a combined epigenetic therapy against human mantle cell lymphoma cells[J]. Clin Cancer Res, 2012, 18(22): 6227
[10].[10]Xie Zhi GANG, Bi Chong LEI, Cheong L L, et al. Determinants of sensitivity to DZNep induced apoptosis in multiple myeloma cells[J]. PLoS One, 2011, 6(6): e21583
[11].[11]Kikuchi J, Takashina T, Kinoshita I, et al. Epigenetic therapy with 3-deazaneplanocin A, an inhibitor of the histone methyltransferase EZH2, inhibits growth of non-small cell lung Cancer cells[J]. Lung Cancer, 2012, 78(2): 138
[12].[12]Fiskus W, Wang Yongchao, Sreekumar A, et al. Combined epigenetic therapy with the histone methyltransferase EZH2 inhibitor 3-deazaneplanocin A and the histone deacetylase inhibitor panobinostat against human AML cells[J]. Blood, 2009, 114(13): 2733
[13].[13]Cheng Lai LING, Itahana Y, Lei Zheng DENG, et al. TP53 genomic status regulates sensitivity of gastric Cancer cells to the histone methylation inhibitor 3-Deazaneplanocin A (DZNep)[J]. Clin Cancer Res, 2012, 18(15): 4201
[14].[14]Fujiwara T, Saitoh H, Inoue A, et al. 3-Deazaneplanocin A (DZNep), an inhibitor of S-Adenosylmethionine-dependent methyltransferase, promotes erythroid differentiation[J]. J Biol Chem, 2014, 289(12): 8121
[15].[15]Chiba T, Suzuki E, Negishi M, et al. 3-Deazaneplanocin A is a promising therapeutic agent for the eradication of tumor-initiating hepatocellular carcinoma cells[J]. Int J Cancer, 2012, 130(11): 2557
[16].[16]Ougolkov A V, Bilim V N, Billadeau D D. Regulation of pancreatic tumor cell proliferation and chemoresistance by the histone methyltransferase enhancer of zeste homologue 2[J]. Clin Cancer Res, 2008, 14(21): 6790
[17].[17]Zhang Yi, Liu Guang PU, Lin Chang WEI, et al. Silencing the EZH2 gene by RNA interference reverses the drug resistance of human hepatic multidrug-resistant Cancer cells to 5-Fu[J]. Life Sci, 2013, 92(17/19): 896

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备注/Memo

备注/Memo:

基金项目 国家自然科学基金资助项目（81172573）

作者简介? 孔令平（1988-）,女 , 硕士在读， 研究方向：头颈部鳞癌的基础研究; 通信作者：张仑，E-mail：zhanglun@tjmuch.com。

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更新日期/Last Update: 2015-11-26