鸡p15因子抑制MDCC-MSB1细胞的增殖及端粒酶活性

为探讨鸡p15基因的生物学功能,试验构建了鸡p15基因的真核表达载体pcDNA3.1( )-p15,并转染到鸡MDV转化的淋巴细胞系MDCC-MSB1,应用G418筛选掉未转染的细胞,对存活细胞p15蛋白的表达、细胞增殖力、群体倍增时间、细胞周期和端粒酶的活性进行了检测。结果表明,与转染空质粒pcDNA3.1( )细胞相比,转染了p15基因的细胞稳定表达了p15蛋白;细胞的增殖受到了抑制,抑制率达45%～74%;群体倍增时间从27h延长至416h;流式细胞仪分析细胞周期发现,p15蛋白引起了细胞多停滞于G0/G1期,S和G2/M期细胞比例下降;端粒酶活性受到抑制。

Overexpression of chicken p 1 5 can suppress the proliferation and telomerase activity of MDCC-MSB1

To study the biofunction of p15,recombinant eukaryon expression vector pcDNA3.1( )-p15 was constructed and transfected into MDCC-MSB1,a chicken lymphoblastoid cells line transformed by Marek's disease virus.The untransfected cells were eliminated by G418.The transfected cells were collected to test the expression of p15 protein,the proliferation ability,population doubling time,cell cycle phases and telomerase activity.In contrast to the cells transfected with pcDNA3.1( ),the cells transfected with pcDNA3.1( )-p15 stably expressed p15 protein even until 144 h after transfection.p15 protein induce MDCC-MSB1 cell proliferation inhibition with the inhibition ratio to be ranged from 45 to 74 percent.Population doubling time of cells transfected with pcDNA3.1( )-p15 were delayed from 27 h to 416 h.Cell cycle phases was analyzed by Flow Cytometry.p15 protein arrested MDCC-MSB1 in G0/G1 phase,decreased the ratio of cells in S and G2/M phase and strongly inhibited telomerase activity.These results suggest that the expressed chicken p15 protein can suppress the proliferation and telomerase activity of MDCC-MSB1.