PTEN在肝癌中表达缺失的意义及其体外转染对人肝癌细胞系的作用

The loss of expression and significance of PTEN in hepatocelluar carcinoma and the effects of PTEN gene transfer in human hepatocelluar carcinoma Cell Line hHCC

目的探讨PTEN表达与肝细胞癌发生、发展的关系,观察体外转染外源性PTEN对肝癌细胞的生长、细胞周期和超微结构的影响。方法检测PTEN在肝癌组织和肝癌细胞系中的表达缺失情况,以观察该基因与肝癌病理分级、转移和预后的关系。将含PTEN的反转录病毒载体及空载体转入人肝癌细胞系HHCC,观察转染前后细胞的生长、超微结构、细胞周期及裸鼠致瘤能力改变情况。结果肝癌中PTEN表达缺失率为30.16%(19/63),显著高于正常肝组织0(0/8)和肝硬化组织7.15%(1/14)。随着恶性程度的增高,PTEN表达缺失率随之增加。PTEN表达缺失与转移及预后显著相关。PTEN在人肝癌细胞系hHCC中也不表达。将抑癌基因PTEN转入后,hH-CC细胞生长明显受到抑制,细胞超微结构失去部分恶性表型特征,细胞周期由G1→S0期受抑制,并出现凋亡峰,体外成瘤能力下降。结论抑癌基因PTEN失表达多见于肝细胞肝癌进展期,并与肝细胞肝癌分化程度、是否转移以及临床预后相关;体外PTEN的转入可以明显地抑制HHCC生长、体外成瘤能力。

Objective To investigate the relationship between the loss of expression of PTEN and the HCC development and malignancy and to investigate the effects of PTEN gene transfer in human hepatocelluar carcinoma Cell Line hHCC.Methods We examined the expression of PTEN protein on archival HCC materials and hepatocelluar carcinoma Cell Line.Then transferred PTEN gene into PTEN-deficient HCC cell line hHCC to study cell growth,segregation,apoptosis,cell cycle,ultrastructure alteration in vitro and nude mice tumor genesis ability in vivo.Results PTEN protein was detected in 19 out of 63 cases of HCC,much higher than that of normal liver tissues and cirrhotic liver tissues.The negative rate has a statistical significant relationship with the difference,metastasis and prognosis.After PTEN was transfected into HHCC and expressed,cells growth rate obviously slows down,FCM shows a apoptosis peak and an obvious G_1 block in cell cycle,EMS shows cells lose some malignant features in ultrastructure,nude mice tumor genesis ability in vivo is weakened.Conclusion Loss of tumor suppressor PTEN protein expression usually occurs in hHCC pathologic progression and is correlated with differentiation,metastasis and prognosis.PTEN can inhibit hHCC growth,adhesion in vitro and tumor genesis ability in vivo obviously.