Abstract:

BACKGROUND: N-Myc downstream regulated gene 2 (NDRG2) is a new tumor supressor gene, and it can either improve anti-tumor effect by classic pathway or monitor carcinogenesis. But there is yet no report addressing NDRG2 role in myeloma occurrence.
OBJECTIVE: Using pBaba-puro retroviral vector to construct recombinant vector which carrying NDRG2 gene and pack virus, then to screen U266 cells stably expressing NDRG2.
METHODS: Primers were designed and synthesized. RNA was extracted from U266 cells, and then applied to reverse transcription for the template and latter PCR amplification. After PCR amplification and digested by Taq I and BamHI, agarose gel electrophoresis was used to detect the correct fragments. We used the vectors to package the retrovirus and infected the U266 cells. The U266 cells stably expressing NDRG2 were screened and NDRG2 protein was detected by western blot assay.
RESULTS AND CONCLUSION: The vector carrying NDRG2 was successfully constructed. The retrovirus was also packaged and infected U266 cells. The U266 cells stably expressing NDRG2 were screened. In these U266 cells, NDRG2 expression was increased significantly. The use of recombinant retrovirus vector technology can successfully construct a retrovirus carrying the corresponding gene which can be used for the following study on NDRG2 function in human myeloma.

Key words: multiple myeloma, genes, transfection, retroviridae