[Construction and expression of eukaryotic expression vector of a novel apoptosis-related mouse gene, mPNAS-4 and its antitumor activity in vitro].

Autor:	Yuan Z; School of Life Science, Sichuan University, Chengdu 610064, China. yuanzhujyx@gmail.com, Yan F, Zhao XY, Deng HX, Li J
Jazyk:	čínština
Zdroj:	Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology [Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi] 2007 Dec; Vol. 23 (12), pp. 1140-3.
Abstrakt:	Aim: To construct eukaryotic vector of a novel apoptosis-related gene from mouse (mPNAS-4) and overexpress it in mouse Lewis lung carcinoma(LL2) cell line. To explore the apoptosis of the tumor cells induced by overexpressed mPNAS-4 in LL2 cells via transfection. Methods: RT-PCR was applied to amplify the encoding region of mPNAS-4 gene from the mouse liver. The cDNA was cloned into the eukaryotic expression vector pcDNA3.1(+) and the resulting recombinant expression plasmid pcDNA3.1(+)-mPNAS-4 was then transfected into mouse Lewis lung carcinoma (LL2) cells through Lipofectamine 2000. Overexpression of mPNAS-4 in the transfected cells and proliferation of the transfected cells was detected by RT-PCR and MTT assay,respectively, while the apoptosis was analysed by FCM and DNA ladder. Results: The recombinant eukaryotic expression vector pcDNA3.1(+)-mPNAS-4 was successfully constructed.The expression of mPNAS-4 at the mRNA level in LL2 cells transfected with pcDNA3.1(+)-mPNAS-4 was up-regulated significantly. The proliferation of LL2 cells was inhibited. Conclusion: Overexpression of mPNAS-4 may have apoptotic effects and therefore inhibit the proliferation of LL2 cells.
Databáze:	MEDLINE
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