[1]生秀梅)△,王正新).ErbB3/Her3基因重组慢病毒的构建及应用*[J].郑州大学学报(医学版),2016,(05):572-575.
 SHENG Xiumei),WANG Zhengxin).Construction and application of recombinant ErbB3/Her3 lentiviral[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2016,(05):572-575.
点击复制

ErbB3/Her3基因重组慢病毒的构建及应用*()
分享到:

《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2016年05期
页码:
572-575
栏目:
论著
出版日期:
2016-08-20

文章信息/Info

Title:
Construction and application of recombinant ErbB3/Her3 lentiviral
作者:
生秀梅12)△ 王正新2)
1)江苏大学医学院生物化学教研室 镇江 212013;2)克拉克亚特兰大大学生物科学系肿瘤研究与治疗发展中心 亚特兰大 乔治亚州 30314
Author(s):
SHENG Xiumei12)WANG Zhengxin2)
1)Department of Biochemistry, School of Medicine, Jiangsu University, Zhenjiang 212013;2)The Center for Cancer Research and Therapeutic Development, Department of Biological Sciences, Clark Atlanta University, Atlanta, GA 30314
关键词:
慢病毒 Her3 p44
Keywords:
lentiviral Her3 p44
分类号:
R734.2
摘要:
目的:构建慢病毒Lenti-Her3,探索ErbB3/Her3在p44通路中的作用。方法:将ErbB3/Her3基因的两个片段Her3-1和Her3-2分段克隆至pCDH-CMV-MCS-EF1-Puro,构建慢病毒载体pCDH-Her3,然后将其包装成慢病毒Lenti-Her3; 用Lenti-Her3感染经p44-shRNA沉默的A549细胞,细胞计数法检测细胞的生长情况,Western blot法检测 P44、Her3蛋白的表达。结果:转染p44-shRNA的A549细胞P44蛋白不表达,Her3蛋白表达显著降低,细胞计数显著降低; Lenti-Her3感染后,p44沉默的A549细胞Her3蛋白表达显著增强,细胞计数有所升高(P<0.05),但未达到正常水平。结论:成功构建了重组慢病毒Lenti-Her3; p44可部分通过Her3促进肺癌A549细胞的生长,可能成为肺癌药物研究的新靶点。
Abstract:
Aim: To construct lentiviral Lenti-Her3 and identify ErbB3/Her3 function in p44 pathway.Methods: Her3-1 and Her3-2 fragments of ErbB3/Her3 gene were cloned in turn into pCDH-CMV-MCS-EF1-Puro to generate pCDH-Her3,and then was packaged to obtain Lenti-Her3. Lenti-Her3 was infected in A549 cells which had been transfected with p44-shRNA, and then expressions of P44 and Her3 proteins were detected by Western blot, cell growth was detected through cell counting.Results: The A549 cells transfected with p44-shRNA had no expression of P44 protein, low expression of Her3 protein, and low cell number counting. The expression ofHer3 protein in the A549 cells transfected with p44-shRNA and infected with Lenti-Her3 was improved, and cell number counting was increased, but did not reach the normal level.Conclusion: The lentiviral Lenti-Her3 has been successfully constructed; p44 could promote lung cancer cell growth partially by Her3 pathway, which may be a potential therapeutic target for lung cancer.

参考文献/References:

[1] YARDEN Y,SLIWKOWSKI MX.Untangling the ErbB signalling network[J].Nat Rev Mol Cell Biol,2001,2(2):127
[2] Leahy DJ. Structure and function of the epidermal growth factor(EGF/ErbB)family of receptors[J].Adv Protein Chem,2004,68:1
[3] KOLIBABA KS,DRUKER BJ.Protein tyrosine kinases and cancer[J].Biochim Biophys Acta,1997,1333(3):F217
[4] AURISICCHIO L,MARRA E,ROSCILLI GA,et al.The promise of anti-ErbB3 monoclonals as new cancer therapeutics[J].Oncotarget,2012,3(8):744
[5] YI ES,HARCLERODE D,GONDO M,et al.High c-erbB-3 protein expression is associated with shorter survival in advanced non-small cell lung carcinomas[J].Mod Pathol,1997,10(2):142
[6] SITHANANDAM G,FORNWALD LW,FIELDS JR,et al.Anti-tumor efficacy of naked siRNAs for ERBB3 or AKT2 against lung adenocarcinoma cell xenografts[J].Int J Cancer,2012,130(2):251
[7] ALIMANDI M,ROMANO A,CURIA MC,et al.Cooperative signaling of ErbB3 and ErbB2 in neoplastic transformation and human mammary carcinomas[J].Oncogene,1995,10(9):1813
[8] ZHOU H,LIU L,LEE K,et al.Lung tumorigenesis associated with erb-B-2 and erb-B-3 overexpression in human erb-B-3 transgenic mice is enhanced by methylnitrosourea[J].Oncogene,2002,21(57):8732
[9] HYNES NE,LANE HA.ERBB receptors and cancer: the complexity of targeted inhibitors[J].Nat Rev Cancer,2005,5(5):341
[10]GU Z,ZHANG F,WANG ZQ,et al.The p44/wdr77-dependent cellular proliferation process during lung development is reactivated in lung cancer[J].Oncogene,2013,32(15):1888
[11]ZHOU L,WU H,LEE P,et al.Roles of the androgen receptor cofactor p44 in the growth of prostate epithelial cells[J].J Mol Endocrinol,2006,37(2):283
[12]李苏霞,张阳,徐建明,等.非小细胞肺癌组织中HER2、HER3蛋白表达及其临床意义[J].肿瘤防治研究,2007,34(10):764
[13]YAMAMOTO T,IKAWA S,AKIYAMA T,et al.Similarity of protein encoded by the human c-erb-B-2 gene to epidermal growth factor receptor[J].Nature,1986,319(650):230
[14]TAKEUCHI K,ITO F.EGF receptor in relation to tumor development: molecular basis of responsiveness of cancer cells to EGFR-targeting tyrosine kinase inhibitors[J].FEBS J,2010,277(2):316
[15]JULIACHS M,CASTILLO-ÁVILA W,VIDAL A,et al.ErbBs inhibition by lapatinib blocks tumor growth in an orthotopic model of human testicular germ cell tumor[J].Int J Cancer,2013,133(1):235
[16]SHI FM,TELESCO SE,LIU YT,et al.ErbB3/HER3 intracellular domain is competent to bind ATP and catalyze autophosphorylation[J].Proc Natl Acad Sci U S A,2010,107(17):7692
[17]SIERKE SL,CHENG K,KIM HH,et al.Biochemical characterization of the protein tyrosine kinase homology domain of the ErbB3(HER3)receptor protein[J].Biochem J,1997,322(Pt 3):757

相似文献/References:

[1]柳 严,张德林,王国良,等.shRNA介导的波形蛋白基因沉默对肝癌细胞迁移、侵袭能力的影响[J].郑州大学学报(医学版),2018,(06):743.[doi:10.13705/j.issn.1671-6825.2018.04.035]
 LIU Yan,ZHANG Delin,WANG Guoliang,et al.Effect of shRNA lentiviral mediated Vimentin gene silencing on migration and invasion of hepatocellular carcinoma cells[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2018,(05):743.[doi:10.13705/j.issn.1671-6825.2018.04.035]

备注/Memo

备注/Memo:
*国家自然科学基金项目 31000046; 江苏大学高级人才启动基金 11JD063; 国家博士后基金面上项目 2015M571702
△女,1978年1月生,博士,副教授,研究方向:肿瘤的发生发展机制及细菌致病机制,E-mail:shengxiumei@ujs.edu.cn
更新日期/Last Update: 2016-08-20