Specific CEA-producing colorectal carcinoma cell killing with recombinant adenoviral vector containing cytosine deaminase gene

	Specific CEA-producing colorectal carcinoma cell killing with recombinant adenoviral vector containing cytosine deaminase gene
	Shen, LZ; Wu, WX; Xu, DH; Zheng, ZC; Liu, XY; Ding, Q; Hua, YB; Yao, K
刊名	WORLD JOURNAL OF GASTROENTEROLOGY
	2002
卷号	8 期号:2 页码:270-275
通讯作者	Wu, WX (reprint author), Nanjing Med Univ, Dept Gen Surg, Affiliated Hosp 1, 300 Guangzhou Rd, Nanjing 210029, Jiangsu Prov, Peoples R China.,
英文摘要	AIM: To kill CEA positive colorectal carcinoma cells specifically using the E coli cytosine deaminase (CD) suicide gene, a new replication-deficient recombinant adenoviral vector was constructed in which CD gene was controlled under CEA promoter and its in vitro cytotoxic effects were evaluated. METHODS: Shuttle plasmid containing CD gene and regulatory sequence of the CEA gene was constructed and recombined with the right ann of adenovirus genome DNA in 293 cell strain. Dot blotting and PCR were used to identify positive plaques. The purification of adenovirus was performed with ultra-concentration in CsCl step gradients and the titration was measured With plaque formation assay, Cytotoxic effects were assayed with MTT method, The fifty percent inhibition concentration ( IC41) of 5-FC was calculated using a curve-fitting parameter. The human colorectal carcinoma cell line. which was CEA-producing. and the CEA-nonproducing Hela cell line were applied in cytological tests. An established recombinant adenovirus vector AdCMVCD, in which the CD gene was controlled under CMV promoter, was used as virus control. Quantitative results were expressed as the mean +/- SD of the mean. Statistical analysis was performed using ANOVA test. RESULTS: The desired recombinant adenovirus vector was named AdCEACD. The results of dot blotting and PCR showed that the recombinant adenovirus contained CEA promoter and CD gene. Virus titer was about 5.0 x 10(14) pfu/L-1 after purification. The CEA-producing Lovo cells were sensitive to 5-FC and had the same cytotoxic effect after infection with AdCEACD and AdCMVCD (The IC50 vaiues of 5-FC in parent Lovo cells. Lovo cells infected with 100 M 0, I AdCEACD and Lovo cells infected with 10 M. O. I AdCMVCD were > 15 000. 216.5 +/- 38.1 and 128.8 +/- 25.4 mumol(.)L(-1) P < 0. 001. respectively), and the cytotoxicity of 5-FC increased accordingly when the m. o. i of adenoviruses were enhanced (The value of IC50 of 5-FC was reduced to 27.9 +/- 4.2 mumol.L-1 in 1000 M. O. I AdCEACD infected Lovo cells and 24.8 +/- 7.1 mumol(.)L(-1) in 100 M. O. I AdCMVCD infected Lovo cells. P < 0.05, P < 0.01, respectively, The CEA-nonproducing Hela cells had no effect after infection with AdCEACD, but Hela cells had the cytotoxic sensitivity to 5-FC after infection with AdCMVCD The IC50 of 5-FC in parent Hele cells and Hela cells infected with AdCMVCD at I o M. 0. 1 was > 15 WO and 214.5 +/- 31.3 mumol(.)L(-1). P < 0.001). AdCEACD/5- FC system also had bystander effect. and the viability was about A) percent when the proportion of transfected cells was only 10 percent, CONCLUSION: The recombinant adenovirus vector AdCEACD has the character of cell type-specific gene delivery, The AdCEACD/5-FC system may become a new, potent and specific approach for the gene therapy of CEA-positive neoplasms, especially colon carcinoma.
学科主题	Gastroenterology & Hepatology
类目[WOS]	Gastroenterology & Hepatology
关键词[WOS]	RETROVIRAL VECTORS ; THERAPY ; EXPRESSION ; SYSTEM
收录类别	SCI
语种	英语
WOS记录号	WOS:000175104600017
内容类型	期刊论文
版本	出版稿
源URL	[http://202.127.25.143/handle/331003/2442]
专题	上海生化细胞研究所_上海生科院生化细胞研究所
推荐引用方式 GB/T 7714	Shen, LZ,Wu, WX,Xu, DH,et al. Specific CEA-producing colorectal carcinoma cell killing with recombinant adenoviral vector containing cytosine deaminase gene[J]. WORLD JOURNAL OF GASTROENTEROLOGY,2002,8(2):270-275.
APA	Shen, LZ.,Wu, WX.,Xu, DH.,Zheng, ZC.,Liu, XY.,...&Yao, K.(2002).Specific CEA-producing colorectal carcinoma cell killing with recombinant adenoviral vector containing cytosine deaminase gene.WORLD JOURNAL OF GASTROENTEROLOGY,8(2),270-275.
MLA	Shen, LZ,et al."Specific CEA-producing colorectal carcinoma cell killing with recombinant adenoviral vector containing cytosine deaminase gene".WORLD JOURNAL OF GASTROENTEROLOGY 8.2(2002):270-275.