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[Construction of a RU486 inducible recombinant adenoviral vector carrying murine interleukin-12 gene and experimental treatment of colonic carcinoma].
Zhonghua Yi Xue Za Zhi. 2009 May 26; 89(20):1372-6.ZY

Abstract

OBJECTIVE

To construct a RU486 inducible recombinant adenovirus of murine IL-12 protein and study its effect and safety on colonic cancer.

METHODS

The replication-defective recombinant adenovirus were produced after cotransfection of shutter vector pDC-RUmIL-12 and adenovirus DNA helper plasmid pBHGloxDeltaE1, 3Cre into HEK293 cells. The recombined adenovirus was purified by CsCl density gradient centrifugation and its titer was determined by end point dilution assay. Expression of this regulatable recombinant adenovirus vector in infected C26 colonic carcinoma cells was tested by ELISA kit in vitro. The tumor model was established by hypodermic inoculation of C26 cells. Sixty tumor-bearing mice were randomly divided into 4 groups: Ad-buffer group; Ad-RUmIL-12 group; Ad-RUmIL-12 + RU486 group and Ad-mIL-12 group, and the treatment effects and side effects were evaluated.

RESULTS

The adenoviral vector containing murine IL-12 gene was identify by PCR. The viral titer of Ad-RUmIL-12 was 4.62 x 10(10) pfu/ml. The expression of IL-12 protein was induced by the RU486 and the highest expression (516 +/- 43) pg/ml whereas no significant IL-12 protein was detected without inducer or getting rid of the inducer [(38 +/- 3) pg/ml and (42 +/- 5) pg/ml respectively]. The tumor size increased rapidly in group Ad-buffer and group Ad-RUmIL-12 (P > 0.05). Administration of Ad-RUmIL-12 + RU486 and Ad-mIL-12 were showed to delay markedly the growth of transplanted C26 tumor (P > 0.05). Significantly necrosis was observed in both Ad-mIL-12 and Ad-RUmIL-12 + RU486 experimental groups, but the level of the serum alanine transaminase and the rate of side effect was higher in Ad-mIL-12 group (4/15 and 10/15 respectively, P < 0.05).

CONCLUSION

A RU486 regulatable recombinant adenoviral vector containing IL-12 gene was successfully constructed. The expression of vector Ad-RUmIL-12, regulated by inducer RU486 in vivo, can obviously improve safety in tumor treatment and provide a good primer for further researches on in vivo gene therapy.

Authors+Show Affiliations

Department of General Surgery, No.81 Hospital of PLA, Nanjing 210002, China.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

English Abstract
Journal Article
Research Support, Non-U.S. Gov't

Language

chi

PubMed ID

19671324

Citation

Chen, Jian, et al. "[Construction of a RU486 Inducible Recombinant Adenoviral Vector Carrying Murine Interleukin-12 Gene and Experimental Treatment of Colonic Carcinoma]." Zhonghua Yi Xue Za Zhi, vol. 89, no. 20, 2009, pp. 1372-6.
Chen J, Liu XS, Wang F, et al. [Construction of a RU486 inducible recombinant adenoviral vector carrying murine interleukin-12 gene and experimental treatment of colonic carcinoma]. Zhonghua Yi Xue Za Zhi. 2009;89(20):1372-6.
Chen, J., Liu, X. S., Wang, F., Xue, X. C., Fang, G. E., Su, C. Q., & Qian, Q. J. (2009). [Construction of a RU486 inducible recombinant adenoviral vector carrying murine interleukin-12 gene and experimental treatment of colonic carcinoma]. Zhonghua Yi Xue Za Zhi, 89(20), 1372-6.
Chen J, et al. [Construction of a RU486 Inducible Recombinant Adenoviral Vector Carrying Murine Interleukin-12 Gene and Experimental Treatment of Colonic Carcinoma]. Zhonghua Yi Xue Za Zhi. 2009 May 26;89(20):1372-6. PubMed PMID: 19671324.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - [Construction of a RU486 inducible recombinant adenoviral vector carrying murine interleukin-12 gene and experimental treatment of colonic carcinoma]. AU - Chen,Jian, AU - Liu,Xu-shun, AU - Wang,Feng, AU - Xue,Xu-chao, AU - Fang,Guo-en, AU - Su,Chang-qing, AU - Qian,Qi-jun, PY - 2009/8/13/entrez PY - 2009/8/13/pubmed PY - 2009/11/18/medline SP - 1372 EP - 6 JF - Zhonghua yi xue za zhi JO - Zhonghua Yi Xue Za Zhi VL - 89 IS - 20 N2 - OBJECTIVE: To construct a RU486 inducible recombinant adenovirus of murine IL-12 protein and study its effect and safety on colonic cancer. METHODS: The replication-defective recombinant adenovirus were produced after cotransfection of shutter vector pDC-RUmIL-12 and adenovirus DNA helper plasmid pBHGloxDeltaE1, 3Cre into HEK293 cells. The recombined adenovirus was purified by CsCl density gradient centrifugation and its titer was determined by end point dilution assay. Expression of this regulatable recombinant adenovirus vector in infected C26 colonic carcinoma cells was tested by ELISA kit in vitro. The tumor model was established by hypodermic inoculation of C26 cells. Sixty tumor-bearing mice were randomly divided into 4 groups: Ad-buffer group; Ad-RUmIL-12 group; Ad-RUmIL-12 + RU486 group and Ad-mIL-12 group, and the treatment effects and side effects were evaluated. RESULTS: The adenoviral vector containing murine IL-12 gene was identify by PCR. The viral titer of Ad-RUmIL-12 was 4.62 x 10(10) pfu/ml. The expression of IL-12 protein was induced by the RU486 and the highest expression (516 +/- 43) pg/ml whereas no significant IL-12 protein was detected without inducer or getting rid of the inducer [(38 +/- 3) pg/ml and (42 +/- 5) pg/ml respectively]. The tumor size increased rapidly in group Ad-buffer and group Ad-RUmIL-12 (P > 0.05). Administration of Ad-RUmIL-12 + RU486 and Ad-mIL-12 were showed to delay markedly the growth of transplanted C26 tumor (P > 0.05). Significantly necrosis was observed in both Ad-mIL-12 and Ad-RUmIL-12 + RU486 experimental groups, but the level of the serum alanine transaminase and the rate of side effect was higher in Ad-mIL-12 group (4/15 and 10/15 respectively, P < 0.05). CONCLUSION: A RU486 regulatable recombinant adenoviral vector containing IL-12 gene was successfully constructed. The expression of vector Ad-RUmIL-12, regulated by inducer RU486 in vivo, can obviously improve safety in tumor treatment and provide a good primer for further researches on in vivo gene therapy. SN - 0376-2491 UR - https://www.unboundmedicine.com/medline/citation/19671324/[Construction_of_a_RU486_inducible_recombinant_adenoviral_vector_carrying_murine_interleukin_12_gene_and_experimental_treatment_of_colonic_carcinoma]_ L2 - http://journal.yiigle.com/LinkIn.do?linkin_type=pubmed&amp;issn=0376-2491&amp;year=2009&amp;vol=89&amp;issue=20&amp;fpage=1372 DB - PRIME DP - Unbound Medicine ER -