过表达的p27^(KIP1)基因诱导HCC-9204细胞凋亡

Apoptosis of HCC-9204 cells induced by over-expressed p27^(KIP1)

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 regulative effects of p27 KIP1 on apoptosis of hepatocellular carcinoma (HCC) cells. Methods We used an inducible expression system of pMD neo allowing controlled expression of protein upon addition of Zinc as an external inducer. The whole length of p27 KIP1 cDNA was transfected into human HCC cell line HCC 920 by the method of lipofectin transfection. Expression of in p27 KIP1 protein or mRNA level was analyzed by immunohistochemistry and RT PCR, respectively. Flow cytometry, TUNEL and electron microscopy were applied to assess the effects of p27 KIP1 on apoptosis. Results Expression of p27 KIP1 in protein or mRNA significantly increased in HCC 9204 cells transfected with p27 KIP1 . Prolonged p27 KIP1 expression induced apoptotic cell death was reflected by pre G 1 peak in the histogram of FACS, which was also confirmed by TUNEL assay and electron microscopy. Conclusions p27 KIP1 may cause apoptosis in HCC.