p27^(KIP1)基因过表达诱导HCC-9204细胞在G_1期停滞并导致细胞凋亡

Overexpression of transfected p27^(KIP1) induces cell cycle arrest in G_1 phase and subsequent apoptosis of HCC-9204 cell line

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

ObjectiveTo investigate the effect of p27 KIP1 transfection on cell cycle and apoptosis of hepatocellular carcinoma cells(HCC). MethodsWe used an inducible expression system pMD neo, which allowed controlled expression of protein upon addition of zinc as an external inducer. p27 KIP1 cDNA was transfected into human HCC 9204 cell line. Expression of p27 KIP1 was analyzed and cell growth was observed. ResultsExpression of p27 KIP1 in protein and mRNA increased significantly in HCC 9204 cell line transfected with p27 KIP1 . The cell growth reduced by 35%, p27 KIP1 over expression caused cell growth arrest at G 1 by 35% ( P =0 000). Apoptotic cell index significantly increased ( P =0 000).Conclusionp27 KIP1 may cause cell cycle arrest in G 1 phase and subsequently lead to apoptosis.