JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES) ›› 2014, Vol. 52 ›› Issue (11): 11-15.doi: 10.6040/j.issn.1671-7554.0.2014.263

Previous Articles     Next Articles

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)

206

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

CLC Number: 

  • 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] TU Yan, LI Zhenjiang, TANG Aiping, FEI Yan, LI Huihui, LI Jian, HE Wenfeng. RNAi-mediated silencing of CD147 inhibits the proliferation and invasion of leukemic cells SHI-1 [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2014, 52(9): 1-5.
[2] HUANG Jing, XU Ying, LIU Yi, XIN Wei, ZHAO Xu, LIU Lei, WAN Qiang. Effects of down-regulating of acetyl CoA carboxylase 2 on intracellular lipid accumulation and epithelial mesenchymal transition in high glucose cultured human renal proximal tubular epithelial cells [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2014, 52(7): 16-21.
[3] DONG Zhaojing, SHANG Qianwen, GUO Chun, ZHANG Lining, WANG Qun . Effect of PDCD4 siRNA on the lipid accumulation in macrophages#br# [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2014, 52(4): 18-21.
[4] Lv Yan-feng1, HAN Bing-bing2, YU Hua-long1, WANG Jian-xin1. Construction and effect of shRNA vector silencing EZH2 expression [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2013, 51(9): 31-34.
[5] ZHU Shou-rong1， ZHANG Rui2， CAO Yong-qian1， FENG Zhang1， WANG Yi-bing1. The effect of the survivin-siRNA targeting lentivirus vector on human malignant melanoma cell line A375 [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2013, 51(5): 54-57.
[6] XU Yang-yang1,2， JIANG Zheng1,2， ZHOU Wei3， JIANG Yu-quan1, 2， LI Xin-gang1, 2. Construction of a lentiviral vector carrying shRNA against HIF-1α and generation of a GL261 cell line stably transfected with this vector [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2013, 51(2): 12-16.
[7] XU Wei, CHANG Hong, ZHAI Yun-peng. Construction of Livin shRNA eukaryotic expression vector and its influence on chemotherapeutic sensitivity of HepG2 cells [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2013, 51(12): 20-24.
[8] LIU Qing-liang1, MU Xiao-yan1, WANG-Jing2, CHI Xiang-yu2, ZHANG Min3, MA Wei-xia3. Inhibitory effects of RNA interference and Erlotinib by blocking epidermal
growth factor receptor pathway on the proliferation of A549 cells [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2012, 50(9): 11-.
[9] MENG You-sheng, WANG Xiu-wen, WANG Ya-wei, TIAN Tian-tian. Effect of bFGF-siRNA-mediated gene silencing on proliferation and
apoptosis in the lung cancer cell line A549 [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2012, 50(3): 50-54.
[10] ZHANG Rui-yu, L Zhi-mei, LIU Yi, ZHANG Peng-ju, WAN Qiang, WANG Rong. The mRNA-stabilizing factor Human-Antigen R regulates renal cyclinD1
post-transcriptional expression in human mesangial cells [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2012, 50(2): 47-52.
[11] LI Ping, LIANG Xiao-hong, JI Yong-li, DONG Liang, GAO Yang, CUI Wen-Chao. Therapy effect of 5-FU palliative implanted through puncturing along
cardiac tangent direction in 81 patients with malignant pericardial effusion [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2012, 50(12): 77-.
[12] TONG Shu-qing1, XIAO Dong-jie1, WANG Yun-shan1, JIA Yan-fei1, ZHENG Yan1, MA Xiao-li1, DAI Xin1, SANG Tan2. Construction of STAT3-siRNA expression vector and its effect on proliferation of stomach cancer cell line HGC-27 [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2012, 50(1): 76-80.
[13] ZHANG Jing-jing, MA Dao-xin, KONG Hai-li, WANG Hui-jun, SUN Yuan-xin， LIU Chuan-fang. AML1/ETO siRNA enhances the sensitivity of Kasumi-1 cells to  histone acetylation enzyme inhibitors [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2011, 49(7): 68-73.
[14] WANG Ze-jun1, XU Cheng-wei1, MA Xiao-li2, ZHENG Yan2, JIA Yan-fei2, XIAO Dong-jie2, WANG Yun-shan2 . Effect of CPB2 gene silencing on TAFI expression and   metastasis in breast cancer cells [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2011, 49(4): 57-.
[15] SONG Ai-qin1,2, JU Xiu-li1, SUN Li-rong2, WANG Ling-zhen2, LI Xiao-ling2, YU Hong-sheng3. Suppression of c-myc expression by lentiviral vector-mediated- small interfering RNA in Jiyoye cells [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2011, 49(4): 80-84.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
Copyright 2018 © Editorial office of Journal of Shandong University (Health Sciences)
Supported by Beijing Magtech Co.Ltd