您的位置：山东大学 -> 科技期刊社 -> 《山东大学学报（医学版）》

山东大学学报（医学版） ›› 2014, Vol. 52 ›› Issue (11): 11-15.doi: 10.6040/j.issn.1671-7554.0.2014.263

• 基础医学 • 上一篇    下一篇

靶向EZH2的shRNA表达对SW480细胞的抑制作用及5-Fu对其影响的研究

吕艳锋1,2, 姜华2, 王建新2, 张成博3   

  1. 1. 山东中医药大学, 山东 济南 250014;
    2. 山东大学第二医院肛肠外科, 山东 济南 250033;
    3. 山东中医药大学文献研究所, 山东 济南 250014
  • 收稿日期:2014-04-27 发布日期:2014-11-10
  • 通讯作者: 张成博.E-mail:youtow@sina.com E-mail:youtow@sina.com
  • 基金资助:
    山东省自然科学基金(ZR2011HQ049);济南市科技明星计划(201004055,20090208)

Inhibitive effect of EZH2-specific shRNA on SW480 cells and the impact of 5-FU

LV Yanfeng1,2, JIANG Hua2, WANG Jianxin2, ZHANG Chengbo3   

  1. 1. Shandong University of Traditional Chinese Medicine, Jinan 250014, Shandong, China;
    2. Department of Anoproctology, Second Hospital of Shandong University, Jinan 250033, Shandong, China;
    3. Literature Research Institute, Shangdong University of Traditional Chinese Medicine, Jinan 250014, Shandong, China
  • Received:2014-04-27 Published:2014-11-10

PDF (PC)

193

摘要： 目的 研究EZH2特异性shRNA对人结肠癌细胞株SW480细胞的抑制作用和化疗药物5-FU对其干预作用的影响.方法 体外将EZH2特异性shRNA表达载体转染SW480细胞,RT-PCR和Western blotting检测细胞EZH2 mRNA及蛋白的表达,MTT法检测细胞增殖活性;将靶向EZH2的shRNA表达的SW480细胞接种于裸鼠,5-FU对其进行干预,RT-PCR检测瘤组织EZH2 mRNA的表达.结果 与阴性对照组相比,基因沉默组EZH2 mRNA表达明显降低(P<0.01),EZH2蛋白表达明显降低(P<0.05);与空白对照组相比,基因沉默组细胞生长受到明显抑制(P<0.05);接种于裸鼠后,与正常组相比,5-FU基因沉默组与基因沉默组瘤组织体积、质量、EZH2 mRNA的表达均明显降低(P<0.01),与基因沉默组相比,5-FU基因沉默组EZH2 mRNA的表达明显降低(P<0.05).结论 靶向EZH2的shRNA表达在体外及体内均可显著抑制SW480细胞EZH2 mRNA 和蛋白表达及细胞生长;5-FU能显著抑制体内EZH2基因沉默的SW480细胞EZH2 mRNA的表达.

关键词: EZH2基因, RNA干扰, SW480细胞, 5-Fu

Abstract: Objective To investigate the inhibitive effect of EZH2-specific shRNA on human colon cancer cell line SW480 and the impact of 5-FU on this inhibitive effect. Methods SW480 cells were transfected with EZH2-specific shRNA in vitro, then EZH2 mRNA and protein expressions were detected with RT-PCR and Western blotting, and cell proliferation was examined with MTT. The EZH2-specific shRNA expressing SW480 cells were implanted to nude mice and treated with 5-FU. After that, the levels of EZH2 mRNA expressed by tumor tissues were measured with RT-PCR. Results Compared with that of the negative control group, the EZH2 mRNA expression (P<0.01) and protein expression (P<0.05) of the gene-silencing group markedly decreased. The cell growth of the gene-silencing group was inhibited compared with that of the control group (P<0.05). In nude mice, the tumor volume, tumor weight and EZH2 mRNA expression of the 5-FU gene-silencing group and of the gene-silencing group remarkably decreased compared with those of the normal group (P<0.01). The EZH2 mRNA expression of the 5-FU gene-silencing group significantly decreased compared with that of the gene-silencing group (P<0.05). Conclusion The EZH2-specific shRNAplays a significant inhibitive role on EZH2 mRNA and protein expressions in SW480 cells both in vitro and in vivo. 5-FU can significantly inhibit the EZH2 mRNA expression of EZH2-gene-silencing SW480 cells in vivo.

Key words: EZH2 gene, SW480 cells, RNA interference, 5-Fu

中图分类号: 

  • R735.3
[1] 王建新, 吕艳锋, 韩冰冰, 等. EZH2在结直肠癌组织中的表达[J]. 中国现代普通外科进展, 2011, 14(6): 421-424. WANG Jianxin, LV Yanfeng, HAN Bingbing, et al. Expression of EZH2 in colorectal cancer tissue[J]. Chin J Curr Adv Gen Surg, 2011, 14(6): 421-424.
[2] 吕艳锋, 韩冰冰, 禹化龙, 等. 抑制EZH2基因表达的shRNA载体的构建及其作用[J].山东大学学报:医学版, 2013, 51(9): 31-34. LV Yanfeng, HAN Bingbing, YU Hualong, et al. Construction and effect of shRNA vector silencing EZH2 expression[J]. Journal of Shandong University: Health Sciences, 2013, 51(9): 31-34.
[3] Bracken A P, Pasini D, Capra M, et al. EZH2 is downstream of the pRB-E2F pathway, essential for proliferation and amplified in cancer[J]. EMBO J, 2003, 22(20): 5323-5335.
[4] Tang X, Milyavsky M, Shats I, et al. Activated p53 suppresses the histone methyltransferase EZH2 gene[J]. Oncogene, 2004, 23(34): 5759-5769.
[5] Mimori K, Ogawa K, Okamoto M, et al.Clinical significance of enhancer of zeste homolog 2 expression in colorectal cancer cases[J]. Eur J Surg Oncol, 2005, 31(4): 376-380.
[6] Hanson R D, Hess J L, Yu B D, et al. Mammalian Trithorax and Polycomb-group homologues are antagonistic regulators of homeotic development[J]. Proc Natl Acad Sci USA, 1999, 96(25): 14372-14377.
[7] Jacobs J J, van Lohuizen M.Polycomb repression: from cellular memory to cellular proliferation and cancer[J]. Biochim Biophys Acta, 2002, 1602(2): 151-161.
[8] Tonini T, Bagella L, D'Andrilli G, et al. Ezh2 reduces the ability of HDAC1-dependent pRb2/p130 transcriptional repression of cyclin A[J]. Oncogene, 2004, 23(28): 4930-4937.
[9] Zeidler M, Varambally S, Cao Q, et al. The polycomb group protein EZH2 impairs DNA repair in breast epithelial cells[J]. Neoplasia, 2005, 7(11): 1011-1019.
[10] Lhr J M. Medical treatment of pancreatic cancer[J]. Expert Rev Anticancer Ther, 2007, 7(4): 533-544.
[11] Tsang D P, Cheng A S. Epigenetic regulation of signaling pathways in cancer: role of the histone methyltransferase EZH2[J]. J Gastroenterol Hepatol, 2011, 26(1): 19-27.
[12] Andrea C, Fausto P, Francesca B K, et al. Which strategy after first-line therapy in advanced colorectal cancer?[J]. World J Gastroenterol, 2014, 20(27): 8921-8927.
[13] Xie L, Zhang Z, Tan Z, et al. MicroRNA-124 inhibits proliferation and induces apoptosis by directly repressing EZH2 in gastric cancer[J]. Mol Cell Biochem, 2014, 392(1-2): 153-159.
[14] Zhang Y, Liu G, Lin C, 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, 20; 92(17-19): 896-902.
[15] Crea F, Fornaro L, Paolicchi E, et al. An EZH2 polymorphism is associated with clinical outcome in metastatic colorectal cancer patients[J]. Ann Oncol, 2012, 23(5): 1207-1213.
[1] 高峰, 康白, 杨洪鸣, 赵廷坤, 曲梅花, 王金红. 皮肤T细胞淋巴瘤相关抗原5在肿瘤细胞株中的表达[J]. 山东大学学报（医学版）, 2015, 53(3): 41-45.
[2] 涂燕, 李振江, 汤爱平, 费妍, 李慧慧, 李剑, 贺文凤. 干扰CD147表达抑制白血病细胞SHI-1增殖及移行[J]. 山东大学学报（医学版）, 2014, 52(9): 1-5.
[3] 黄静, 胥莹, 刘毅, 辛玮, 赵胥, 刘蕾, 完强. 乙酰辅酶A羧化酶2表达下调对高糖培养的人肾小管上皮细胞脂质沉积和间充质转化的影响[J]. 山东大学学报（医学版）, 2014, 52(7): 16-21.
[4] 董兆静，尚倩雯，郭春，张利宁，王群. RNA干扰PDCD4基因表达对巨噬细胞脂质沉积的影响[J]. 山东大学学报（医学版）, 2014, 52(4): 18-21.
[5] 罗剑刚,孙涛,林小雯,李芸,赵菲,苗贵申,傅志俭. 慢病毒介导RNAi沉默大鼠脊髓神经元NF-κB p65基因的观察[J]. 山东大学学报（医学版）, 2014, 52(3): 23-26.
[6] 吕艳锋1，韩冰冰2，禹化龙1，王建新1. 抑制EZH2基因表达的shRNA载体的构建及其作用[J]. 山东大学学报(医学版), 2013, 51(9): 31-34.
[7] 朱守荣1，张芮2，曹永倩1，冯璋1，王一兵1. 慢病毒载体靶向携带survivin-siRNA对恶性黑色素瘤A375细胞的影响[J]. 山东大学学报(医学版), 2013, 51(5): 54-57.
[8] 徐洋洋1, 2,姜政1, 2,周伟3,江玉泉1, 2,李新钢1, 2 . HIF-1α慢病毒干扰载体构建及GL261小鼠胶质瘤稳定沉默细胞系的建立[J]. 山东大学学报(医学版), 2013, 51(2): 12-16.
[9] 徐威，常宏，翟云鹏. Livin shRNA真核表达载体构建及其对HepG2细胞化疗敏感性的影响[J]. 山东大学学报(医学版), 2013, 51(12): 20-24.
[10] 刘庆亮1，牟晓燕1，王静2，迟翔宇2，张敏3，马卫霞3. RNA干扰及厄洛替尼阻断EGFR信号途径对A549细胞抗增殖的影响[J]. 山东大学学报(医学版), 2012, 50(9): 11-.
[11] 孟又胜,王秀问,王亚伟,田甜甜. siRNA干扰沉默bFGF基因对肺腺癌A549增殖和凋亡的影响[J]. 山东大学学报(医学版), 2012, 50(3): 50-54.
[12] 李萍，梁晓红，纪永利，董亮，高洋，崔文超. 沿心脏切线方向穿刺植入5-FU缓释粒子治疗恶性心包积液81例疗效分析[J]. 山东大学学报(医学版), 2012, 50(12): 77-.
[13] 童书青1，肖东杰1，汪运山1，郏雁飞1，郑燕1，马晓丽1，戴新1，桑坦2. STAT3-siRN表达载体的构建及其对胃癌细胞株HGC-27细胞增殖的影响[J]. 山东大学学报(医学版), 2012, 50(1): 76-80.
[14] 张晶晶，马道新，孔海丽，王慧君，孙元欣，刘传方. 靶向AML1/ETO的siRNA增强Kasumi-1细胞对组蛋白去乙酰化酶抑制剂的敏感性[J]. 山东大学学报(医学版), 2011, 49(7): 68-73.
[15] 王泽筠1，徐成伟1，马晓丽2，郑燕2，郏雁飞2，肖东杰2，汪运山2. siRNA靶向抑制CPB2基因对乳腺癌细胞TAFI表达及生长和转移的影响[J]. 山东大学学报(医学版), 2011, 49(4): 57-.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
版权所有 © 2018 《山东大学学报(医学版)》编辑部　
地址:山东大学科技期刊社（济南市历城区山大南路27号山东大学中心校区明德楼B座721室） 电话: 0531-88366918 E-mail: xbyxb@sdu.edu.cn
本系统由北京玛格泰克科技发展有限公司设计开发