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

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.