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目的研究p2 7KIP1基因对肝癌细胞的细胞周期和细胞凋亡的调节作用。方法采用一种可诱导性真核表达载体pMD neo ,通过外加Zn2 + 诱导目的基因的表达。脂质体转染法将p2 7KIP1全长cDNA转入肝癌细胞系HCC 92 0 4中 ,检测p2 7KIP1基因的表达 ,对细胞增殖的作用及目的基因对细胞的细胞周期和细胞凋亡的影响。结果免疫组织化学及RT PCR显示转染的p2 7KIP1基因有高水平的表达。在外加Zn2 + 4 8h后细胞生长被抑制 35 % ,G1期细胞数由 35 %增加到 76 % ,P =0 0 0 0 ;凋亡指数显著增加 (P =0 0 0 0 )。结论p2 7KIP1基因能够使HCC 92 0 4细胞的G1期延长并导致细胞凋亡
Objective To study the regulatory effect of p2 7KIP1 on cell cycle and apoptosis of hepatoma cells. Methods An inducible eukaryotic expression vector pMD neo was used to induce the expression of the target gene by adding Zn2 +. The p2 7KIP1 full-length cDNA was transfected into hepatocellular carcinoma cell line HCC 92 0 by lipofectamine 2000. The expression of p2 7KIP1 gene was detected by MTT assay. The effect of the target gene on the cell cycle and apoptosis was analyzed. Results Immunohistochemistry and RT-PCR showed that the transfected p2 7KIP1 gene had a high level of expression. The cell growth was inhibited by 35% at 8 h after addition of Zn2 + 4, and the number of cells in G1 phase increased from 35% to 76%, P = 0.0000; the apoptosis index increased significantly (P = 0.0000). Conclusion The p2 7KIP1 gene can prolong the G1 phase of HCC 92 0 4 cells and induce apoptosis