关键词: 鹿茸, RNAi, CGI99基因, 鹿茸干细胞

Abstract: CGI99 of antlerogenic periosteum（AP）cells from Chinese sika deer was interfered using the lentivirus-mediated gene silencing system,and a preliminary study on the gene function on proliferation of AP cells was proceeded. One sequence shRNA targeting CGI99 of sika deer was designed,then reassembled into the lentiviral plasmids pLVTHM. Together with the plasmids pSPAX2 and pMD2.G,recombinant lentivirus was acquired by their co-transfection into HEK 293t cells,and then infected into AP cells. Detecting the down-regulating expression level of CGI99 mRNA in infected AP cells was conducted by RT-PCR,and the effect of silenced CGI99 on AP cell proliferation was assayed by MTT. The results showed that the expression level of CGI99 mRNA in cells that infected with recombinant lentivirus was obviously decreased,the interferential efficiency reached 70.8%;and the curves by MTT assay tended to be consistent. Therefore,we successfully constructed RNAi vector for CGI99 and interfered the expression of CGI99 in AP cells,and preliminarily confirmed that the CGI99 presented insignificant effect on the AP cells’ proliferation.

Key words: deer antler, RNAi, CGI99, antlerogenic periosteum