Nucleostemin specific RNAi influences cell proliferation in HeLa cells in vitro and in vivo / 中国病理生理杂志

AIM: To examine Nucleostemin (NS) expression in tumor cells, and observe the effect of NS specific RNA interference on the cell proliferation in Hela cells. METHODS: Total RNA was extracted from 6 kinds of cultured tumor lines, the NS expression level was measured by RT-PCR and Northern blot. An NS-specific siRNA expression vector was constructed to transfect HeLa cell (NS-siRNA-HeLa), and the proliferation of the cell was observed. RESULTS: NS was highly expressed in 6 kinds of tumor cells. NS expression level in the NS-siRNA-HeLa cells was remarkably reduced, and the percentage of G_0/G_1 cells increased. The neoplasm forming ability in nude mice by the NS-siRNA-HeLa cells was decreased. CONCLUSION: NS is highly expressed among tumor cells. NS-specific siRNA inhibits the entry of the cell cycle into the S phase, and remarkably reduces the proliferation ability of HeLa cells in vitro and in vivo.

Cloning, Sequencing and Expression of Trichinella spiralis p49 Gene / 中国寄生虫学与寄生虫病杂志

Objective To conduct cloning, sequencing and expression of Trichinella spiralis ES antigen p49 gene. Methods RT-PCR was used to amplify the specific gene fragment from the total RNA of Trichinella spirais larvae. The PCR product was ligated to the T-vector and the recombinant plasmid was verified by sequencing. T-p49 and pGE-4T-3 were treated by both BamHI and XhoI. The ligation reaction was catalyzed by T4 DNA ligase. Results The p49 gene was cloned by using RT-PCR. Sequence analysis showed that the p49 gene obtained was consistent to the p49 sequence reported in the database. The expressed protein was shown as a new band at SDS-PAGE. BLAST analysis demonstrated that this p49 gene was 99% identical to the p49 gene reported and to the 43 kDa secreted glycoprotein gene in the database. Conclusion p49 gene from Trichinella spiralis larvae was cloned, sequenced and expressed.