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目的 研究 p2 7KIP1 基因对肝癌细胞凋亡的潜在调节作用。方法 采用一种可诱导性真核表达载体 pMD neo ,通过外加Zn离子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 ,通过免疫组化及RT PCR检测p2 7KIP1 基因在蛋白质和mRNA水平的表达 ;流式细胞仪、TUNEL染色及透射电镜等方法观察目的基因对该细胞凋亡的影响。结果 转染p2 7KIP1 的HCC 92 0 4细胞在mRNA和蛋白质水平均有高水平 p2 7KIP1 的表达。p2 7KIP1 的过表达使G1期前出现一个亚二倍体的凋亡峰 ,占 2 0 % ,并且其调亡指数也显著增加 (P <0 0 1 )。结论 p2 7KIP1 基因能够诱导HCC 92 0 4细胞凋亡。
Objective To investigate the potential regulatory effect of p2 7KIP1 on hepatocellular carcinoma cell apoptosis. Methods An inducible eukaryotic expression vector pMD neo was used to induce the expression of the target gene by Zn ion. The p2 7KIP1 full-length cDNA was transfected into hepatocellular carcinoma cell line HCC 92 0 by lipofectamine. The expression of p2 7KIP1 gene at mRNA and protein level was detected by immunohistochemistry and RT-PCR. Flow cytometry, TUNEL staining, Transmission electron microscopy and other methods to observe the target gene on the apoptosis. Results HCC 92 0 4 cells transfected with p2 7KIP1 expressed high levels of p2 7KIP1 at mRNA and protein levels. Overexpression of p2 7KIP1 caused a sub-diploid apoptotic peak before G1 phase, accounting for 20% of the total, and its apoptosis index also increased significantly (P <0.01). Conclusion The p2 7KIP1 gene can induce apoptosis of HCC 9240 cells.