多发性骨髓瘤恶性相关基因MYEOV2对NIH/3T3细胞生长的影响

The Effect of Multiple Myeloma Tumor-associated Gene MYEOV2 on NIH/3T3 Cells Growth

目的 分析多发性骨髓瘤恶性相关基因MYEOV2对NIH/ 3T3细胞生长的影响。方法 将MYEOV2基因克隆到真核表达载体pcDNA3 1( +)上 ,并将阳性重组子通过脂质体转染入NIH/ 3T3细胞中 ,G418筛选获得转染阳性克隆并用RT -PCR检测MYEOV2基因的表达。运用流式细胞分类法、细胞生长曲线分析MYEOV2基因对NIH/ 3T3细胞生长的影响。结果 成功获得转染pcDNA3 1( +) /MYEOV2的细胞阳性克隆。流式细胞分析显示转染pcDNA3 1( +) /MYEOV2、转染空载体及未转染的NIH/ 3T3细胞的S期细胞分别为 3 0 9%、2 0 1%、16 1% ,前者较后两者S期细胞增多 ,有显著性差异 (P <0 0 5 )。细胞生长曲线显示转染pcDNA3 1( +) /MYEOV2、转染空载体、未转染的NIH/ 3T3细胞的倍增时间分别为 13 6h、2 5 3h、2 7 4h ,前者细胞倍增时间较后两者缩短 ,有显著性差异 (P <0 0 5 )。结论 MYEOV2基因的表达增加促进DNA合成增加 ,细胞增殖加快 。

Objective To analyze the influence of multiple myeloma tumor-associated gene MYEOV2 on NIH/3T3 cells growth. Methods The recombinant plasmid, pcDNA3.1(+)/MYEOV2, was constructed and subsequently transfected into NIH/3T3 cells by liposome. The positive clones of the transfected NIH/3T3 cells were obtained by G418 selection. RT-PCR was used to detecte the expression of MYEOV2 gene in the cells. Fluorescence-activated cell sorting (FACS) and cell grow curve drawing were applied to analyze the influence of MYEOV2 gene on NIH/3T3 cells proliferation. Results The cell clones stably expressing MYEOV2 gene were obtained. FACS analysis showed that the percentage of S phase cell of pcDNA3.1(+)/MYEOV2-transfected cells, pcDNA3.1(+) -transfected cells and non-transfected cells was 30.9%, 20.1% and 16.1%, respectively, and the difference of sphase cell percentage between pcDNA3.1(+)/MYEOV2-transfected cells and the other two groups of cells was significant (P<0.05). The population double time(PDT) of pcDNA3.1(+)/MYEOV2-transfected cells, pcDNA3.1(+)-transfected cells, and non-transfected cells was 13.6, 25.3 and 27.4 hours, respectively, the difference of which between pcDNA3.1(+)/MYEOV2-transfected cells and the other two groups of cells was also significant(P<0.05). Conclusion The expression of MYEOV2 in NIH/3T3 cells had the potential to stimulate DNA synthesis and lead to cell proliferate rapidly. These results suggested that MYEOV2 gene might be involved in the proliferation of multiple myeloma cells.