Effects of NOEY2 transfection on the growth of human breast cancer cell line

Purpose:To clone and construct an eukaryotic expression vector of NOEY2 gene, and to observe the effects of NOEY2 transfection on the growth of human breast cancer cell line MDA MB 231. Methods:The coding region of NOEY2 was obtained with reverse transcription PCR, and then the PCR product was first cloned into pGEM T vector and further directionally subcloned into pcDNA3. Transfection reagent and selective antibiotic were lipofect AMINE and G418 respectively. The expression level of NOEY2 protein was detected by Western Blotting. The growth cures of the transfected and non transfected cells were recorded. The changes in cell cycle were analyzed by flow cytometry. Results:An eukaryotic expression vector of NOEY2 gene was constructed successfully. The cell transfected with NOEY2 showed definite expression of NOEY2, and the controls were negative. The growth of NOEY2 transfected cells was inhibited by 46.3%, compared with the parental cells on the seventh day after seeding. An obvious decrease in S phase and G2/M phase fraction and an increase in the percentage of G0/G1 phase cells were found in NOEY2 transfected cells. Conclusions:NOEY2 Transfection can inhibit the growth rate of MDA MB 231 cells in vitro, probably via the mechanism of G1 arrest. These data support the suggestion that NOEY2 is a tumor suppressive gene, which merits further investigation for its value as a therapy gene.