摘要
目的:研究肝细胞癌(HCC)中端粒酶逆转录酶基因hTERT表达,探讨其在HCC发生、发展及预后复发中的意义.探讨抗端粒酶在肝细胞癌治疗中的意义.方法:采用免疫组化法检测42例HCC组织中hTERT的表达,并对hTERT与临床病理特征的关系进行分析.将反义hTR真核表达载体经脂质体介导转染人肝癌细胞系HepG2,体外培养及接种裸鼠观察其基因转染细胞的细胞周期、超微结构变化及致瘤性.结果:HCC中hTERT基因阳性率分别为71.4%(30/42);hTERT与在正常肝脏组织中阳性表达率0%相比,有显著性差异(P<0.01).hTERT在Ⅰ,Ⅱ和Ⅲ级HCC中的表达率分别为40.0%(4/10),70.0%(14/20),100%(12/12).Ⅰ与Ⅲ级、Ⅱ与Ⅲ级比较差异显著(P<0.05).hTERT阳性表达率与患者复发有显著相关性(P<0.05).形态学观察,转染后HepG2细胞出现典型的凋亡现象.FCM检测发现G1期前出现凋亡峰.在裸鼠皮下的致瘤性明显降低.荷瘤鼠存活时间延长.结论:hTERT表达异常可能与肝脏肿瘤的发生、发展有关.检测hTERT表达可作为预测肝脏肿瘤预后复发的潜在指标.提示HCC是由多基因参与的疾病.转染端粒酶反义RNA能抑制肝癌HepG2细胞的恶性表型,促进其凋亡.
AIM: To examine the expression of human telomerase reverse transcriptase (hTERT) gene in human hepatocel lular cancer (HCC), to investigate its relevance with the carcinogenesis, development and recurrence of HCC, and to explore the potential of antisense RNA of human telemerase (hTR) gene in the treatment of HCC. METHODS: Immunohistochemistry was used to detect the expression of hTERT protein in 42 specimens of HCC. The relationship between hTERT expression and the clinical and pathological characteristics was analyzed.HepG2 cell line was transfected with antisense hTR expression vector (pBBS-hTR) with lipofectin. The transfected cells were cultured in vitro and then inoculated into nude mice. The cell cycle, ultrastructure and tumorigenicity of the transfected cells were examined. RESULTS: The positive rate of hTERT gene expression in HCC was significantly higher (30/42) than that in normal liver tissues (71.4% vs 0%, P <0.01), and the positive rates were also significantly different between HCC with pathological grades Ⅰ (4/10), Ⅱ(14/20) and Ⅲ(12/12) (40.0% vs70.0% vs 100%, respectively, P<0.05). The expression of hTERT was positively correlated to the recurrence of patients (P<0.05). The transfected cells manifested typical apoptotic morphology, and the apoptotic peak appeared before G1 phase of cell cycle. Tumorigenesis of the transfected cells in nude mice was significantly inhibited. The survival time of nude mice inoculated with transfected cells was markedly prolonged, in comparison with that of control mice. CONCLUSION: The aberrant expression of hTERT gene may be related to the pathogenesis and progression of HCC. The highly expressed hTERT gene may be regarded as a marker for the recurrence of HCC. The growth of HepG2 cells can be effectively inhibited and the apoptos(?)s can be promoted by transfection of antisense telomerase RNA.
出处
《世界华人消化杂志》
CAS
北大核心
2005年第2期175-179,共5页
World Chinese Journal of Digestology
