论文部分内容阅读
研究p2 7KIP1基因对胃癌细胞的细胞周期及细胞增殖的影响。采用脂质体转染法将p2 7KIP1全长cDNA转入胃癌细胞系SGC790 1中 ,通过免疫印迹分析以及RNA斑点杂交方法检测p2 7KIP1基因在蛋白质和mRNA水平的表达 ;细胞活力实验及软琼脂集落形成实验显示转染p2 7KIP1基因对细胞增殖的作用 ;流式细胞仪观察目的基因对该细胞的细胞周期的影响。结果显示 ,转染p2 7KIP1的SGC790 1细胞在mRNA和蛋白质水平均有高水平p2 7KIP1的表达 ;细胞活力检测显示在外加Zn离子 4 8h后细胞生长被抑制 4 2 % ;转染p2 7KIP1的SCG790 1细胞的集落形成率较对照组明显减少(P <0 0 1) ;p2 7KIP1的过表达能够显著地增加G1期的细胞数 ,由 33 6 8%增加到6 9 2 9% (P<0 0 1)。研究表明 ,p2 7KIP1基因可抑制SGC790 1细胞由G1期向S期过渡 ,从而抑制细胞增殖。
To investigate the effect of p2 7KIP1 gene on the cell cycle and cell proliferation of gastric cancer cells. The p2 7KIP1 full-length cDNA was transfected into gastric cancer cell line SGC7901 by lipofection method. The expression of p2 7KIP1 gene at protein and mRNA level was detected by Western blotting and RNA dot blot hybridization. Cell viability assay and soft agar colonies The formation of experiments showed that the transfected p2 7KIP1 gene on cell proliferation; flow cytometry to observe the target gene on the cell cycle. The results showed that SGC7901 cells transfected with p2 7KIP1 had a high level of p27KIP1 expression at both mRNA and protein levels. Cell viability assay showed that cell growth was inhibited 42% after 48 h addition of Zn 2+, and SCG790 transfected with p2 7KIP1 1 cells decreased significantly compared with the control group (P <0.01). Overexpression of p2 7KIP1 significantly increased the number of G1 phase cells from 33 6 8% to 6 9 2 9% (P 0 0 1). Studies have shown that, p2 7KIP1 gene can inhibit SGC790 1 cells from the G1 phase to S phase transition, thereby inhibiting cell proliferation.