[1]浦颖艳,方琳,沈权,等.肿瘤特异性溶瘤腺病毒载体的构建及其在肿瘤细胞中特异性的表达[J].南京师大学报(自然科学版),2008,31(02):92-96.
　Pu Yingyan,Fang Lin,Shen Quan,et al.Construction of Tumor-specific Oncolytic Adenovirus Vector and its Primary Study on Selective Expression in Tumor Cells[J].Journal of Nanjing Normal University(Natural Science Edition),2008,31(02):92-96.
点击复制

肿瘤特异性溶瘤腺病毒载体的构建及其在肿瘤细胞中特异性的表达()
分享到：

《南京师大学报（自然科学版）》[ISSN:1001-4616/CN:32-1239/N]

卷:
第31卷
期数:
2008年02期
页码:
92-96
栏目:
生命科学
出版日期:
2008-06-30

文章信息/Info

Title:
Construction of Tumor-specific Oncolytic Adenovirus Vector and its Primary Study on Selective Expression in Tumor Cells
作者:
浦颖艳;方琳;沈权;孙丽君;胡小翠;王艳;曹祥荣;苏长青;
南京师范大学生命科学学院, 江苏省分子医学重点实验室, 江苏南京210046
Author(s):
Pu Yingyan; Fang Lin; Shen Quan; Sun Lijun; Hu Xiaocui; Wang Yan; Cao Xiangrong; Su Changqing
School of Life Science,Nanjing Normal University,The Jiangsu Key Laboratory of Molecular and Medical Biotechnology,Nanjing 210046,China
关键词:
基因治疗;  溶瘤腺病毒;  肿瘤;  端粒酶启动子
Keywords:
gene therapy;  onco ly tic adenov irus;  tum or;  te lom erase prom o ter
分类号:
R73-36
摘要:
为构建一种由人端粒酶逆转录酶启动子( hTERTp) 调控的能够在肿瘤细胞中特异性增殖的新型肿瘤基因治疗溶瘤腺病毒载体系统, 利用基因重组技术将hTERTp插入5型腺病毒E1A基因上游, 构建肿瘤特异性增殖腺病毒AdSU, 使其携带绿色荧光蛋白报告基因. 同时构建增殖缺陷型腺病毒Ad - EGFP 作为对照. 通过 W estern blot分析表明该特异性增殖腺病毒AdSU - EGFP在肿瘤细胞中特异性表达E IA. 而通过荧光显微镜观察两者增殖实验发现, AdSU - EGFP在正常成纤维细胞株BJ及原代子宫成纤维细胞中无增殖, 在肝癌细胞株MHCC 和肺癌细胞株A549中, 则可见病毒增殖并裂解细胞的现象. 由此可认为成功构建了肿瘤基因治疗的特异性溶瘤腺病毒载体系统AdSU, 该系统能够在肿瘤细胞中特异性增殖并表达所携带的外源基因, 这对肿瘤的定向基因治疗有着重要意义.
Abstract:
To develop a nove l gene therapeu tic onco lytic adenov irus in wh ich the hTERT promo ter w as introduced and used to regulate adenov iral E1A gene. A nove l cancer- spec ific rep lication-com pe tent adenov irus nam ed AdSU w as constructed by em ploy ing the human te lom erase reverse transcr iptase ( hTERT) promo ter to dr ive the expression of adenov-i rus E1A gene and by c loning the EGFP repo rter gene into the adenov irus genom e. The non-rep licative adenov irus nam ed Ad-EGFP w as constructed as the con tro l at the sam e tim e. The se lective replication o f AdSU-EGFP in tumo r cells w as investigated by v irus pro liferative test andW estern b lo t assay. By w este rn b lot assay, E1A prote in w as pos itive in cancer ce lls (MH CC, A549) infected w ith AdSU-EGFP, but negative in norm a l ce lls ( BJ, uter ine fibroblast). Under the fluorescent m icroscope, a few norm a l cells expressed EGFP and em itted fluo rescence, and no phagocy tic p lagues appeared from 3- 10 days after infected w ith AdSU-EGFP. H ow ever, when a few cance r cells w ere infected w ith AdSU - EGFP and em itted fluo rescence afte r 3 days, the fluorescence em ission ce lls spread w ithin a large area after 7 days. Sow e can m ake a conc lusion that a nove l cancer-selective gene therapeutic onco lytic adenov irus vec to r sy stem nam ed AdSU, w hich can se lective replicate and express the rapeutic genes in cancer ce lls, was constructed. It is a prom ising system for targeted cancer gene therapy.

参考文献/References:

[ 1] Shay JW, Bacchetti S. A survey o f telom erase activ ity in hum an cancer[ J]. Eur J C ancer, 1997, 33( 5) : 787-791.
[ 2] K im N W, PiatyszekM A, Prow seK R, et a.l Spec ific asso ciation o f hum an te lome rase activ ity w ith immo rtal cells and cancer[J] . Science, 1994, 266( 5193) : 2 011-2 015.
[ 3] Nemuna itis J, Gan ly I, Khuri F, et a.l Se lec tive rep lication and oncolysis in p53 mutant tumo rs w ith ONYX- 015, an E1B- 55kD g ene-de leted adenov irus, in patients w ith adv anced head and neck cancer: a phase II tria l[ J] . CancerRes, 2000,60( 22) : 6 359-6 366.
[ 4] DeW eese T L, Poe lH, Li S, et a .l A phase I tria l o f CV706, a replica tion- competent, PSA se lec tive onco ly tic adenov irus,
for the treatm ent o f lo ca lly recurrent prostate cancer fo llow ing rad ia tion therapy [ J]. C ancer Res, 2001, 61 ( 20): 7 464-7 472.
[ 5] Khur i F R, Nem una itis J, Gan ly I, et a.l A contro lled trial o f intratum ora lONYX - 015, a se lective ly- rep licating adenovirus, in com bination w ith c isp latin and 5- fluorourac il in pa tients w ith recurrent head and neck cancer[ J]. Na tM ed, 2000,6( 8): 879-885.
[ 6] H echt J R, B edford R, Abbruzzese J L, et a .l A Pahse I /II trial of intra tumo ra l endoscopic u ltrasound in jec tion o fONYX-
015 w ith intravenous gem citabine in unresectab le pancreatic carc inom a[ J]. C lin Cancer Res, 2003, 9( 2): 555-561.
[ 7] K irn D, M artuza R L, Zw iebe l J. Rep lication-se lective v irotherapy fo r cancer: bio log ica l princ ip les, r isk m anagem ent and future d irections[ J]. NatM ed, 2001, 7( 7): 781-787.
[ 8] B ischoff J R, K irn D H, W illiam sA, et a .l An adenov irus m utant tha t rep licates selective ly in p53- defic ient hum an tum or ce lls[ J]. Sc ience, 1996, 274( 5286): 373-376.
[ 9] Ro thmann T, H engsterm ann A, Wh itakerN J, et a.l Rep lication o fONYX- 015, a potential anticancer adenov irus, is independent o f p53 status in tum o r ce lls[ J] . J V iro,l 1998, 72( 12): 9 470-9 478.
[ 10] Rodr iguez R, Schuur E R, L im H R, et a.l Prostate attenua ted rep lication competent adenov irus ( ARCA) CN706: a selectivecy totox ic for prostate-spec ific antigen-positive prosta te cancer cells[ J]. Cancer Res, 1997, 57( 13): 2 559-2 563.
[ 11] K im J, Lee B, K im J S, et a .l Antitumo ra l effects o f recomb inan t adenov irus YKL - 1001, conditiona lly rep licating in alpha- fetopro te in-producing human liver cancer cells[ J]. Cancer Lett, 2002, 180( 1): 23-32.
[ 12] Kur ihara T, B rough D E, Kovesd i I, et a.l Selectivity of a replica tion- competent adenov irus for hum an b reast ca rc inoma cells expressing theMUC1 antigen[ J]. J C lin Invest, 2000, 106( 6): 763-771.
[ 13] Su C Q, Sham J, XueH B, e t a.l Potent an titum ora l e fficacy of a nove l replicative adenov irus CNHK300 ta rgeting te lom erase-
po sitive cancer ce lls[ J]. J Cancer Res C lin Onco ,l 2004, 130( 10): 591-603.
[ 14] Kaw ash ima T, Kag aw a S, Naoya K, et a .l Te lome rase- spec ific replication-selective v irothe rapy for hum an cancer[ J]. ClinCance rRes, 2004, 10( Pt1): 285-292.
[ 15] Su C Q, W ang X H, Chen J, e t a.l Antitum or activ ity of an hTERT promo ter-regu la ted tum or-se lec tive oncolytic adenov irus in hum an hepatoce llu lar carc inom a[ J]. W orld J Gastroentrto ,l 2006, 12( 47): 7 613-7 620.

备注/Memo

备注/Memo:
通讯联系人: 曹祥荣, 教授, 博士生导师, 研究方向: 肿瘤发生与基因治疗分子机制、动物染色体进化与基因组学等. E-m ail:cao63@ 126.com
更新日期/Last Update: 2013-05-05