Telomerase activity in human hepatocellular carcinoma: parallel correlation with human telomerase reverse transcriptase (hTERT) mRNA isoform expression but not with cell cycle modulators or c-Myc expression
Telomerase activity in human hepatocellular carcinoma: parallel correlation with human telomerase reverse transcriptase (hTERT) mRNA isoform expression but not with cell cycle modulators or c-Myc expression
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DOI：
10.1053/ejso.2001.1237
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发表时间：
2002-04-01
期刊：
影响因子：
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通讯作者：
Harn, HJ
中科院分区：
文献类型：
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作者：
Chen, CJ;Tsai, NM;Harn, HJ
背景：为了探讨端粒酶可能的调控机制，我们检测了人肝细胞癌（HCC）细胞系（J5.J7）和正常人永生化肝上皮细胞系（Chang-liver）中端粒酶活性（TA）、人端粒酶RNA（hTR）的表达、人端粒酶逆转录酶（hTERT）mRNA亚型和细胞周期调节剂。方法：对细胞系进行化学同步。在 G(1)、G(1)/S、G(2)/M 或 M 阶段。通过基于聚合酶链式反应 (PCR) 的端粒酶重复扩增方案测定来测量 TA。通过逆转录酶-聚合酶链式反应分析hTR和hTERT mRNA水平。采用Western blotting和免疫细胞化学方法检测细胞周期调节剂。结果：J5、J7和Chang-肝细胞系的TA在M期最高。 hTERT mRNA 的表达水平与 HCC 细胞系 M 期检测到的最高 TA 相关。 Chang-liver表达的TA和hTERT mRNA比J5或J7少。 HCC细胞系M期TA和hTERT mRNA异构体表达的升高与细胞周期调节剂和c-Myc的升高没有显着相关性。结论：结果提示TA的调节与hTERT mRNA异构体的表达有关，并且在细胞永生化和肿瘤发生之间的调节是不同的。 (C) 2002 爱思唯尔科学有限公司。
Background: To explore the possible regulatory mechanisms of telomerase, we examined the telomerase activity (TA), expression of human telomerase RNA (hTR), human telomerase reverse transcriptase (hTERT) mRNA isoforms and cell cycle modulators in human hepatocellular carcinoma (HCC) cell lines (J5.J7) and a normal human immortalized hepatic epithelial cell line (Chang-liver).Methods: The cell lines were chemically synchronized in either G(1), G(1)/S, G(2)/M or M phases. TA was measured by polymerase chain reaction (PCR)-based telomerase repeat amplification protocol assay. The hTR and hTERT mRNA levels were analyzed by reverse transcriptase-polymerase chain reaction. Western blotting and immunocytochemistry were used to assay the cell cycle modulators.Results: The TA of J5, J7 and Chang-liver cell lines tested was highest in M phase. The expression level of hTERT mRNA associated with the highest TA detected in the M phase of HCC cell lines. Chang-liver expressed mal less TA and hTERT mRNA than J5 or J7. The elevated TA and expression of hTERT mRNA isoforms in M phase of HCC cell lines did not significantly correlate with that of the cell cycle modulators and c-Myc.Conclusions: The results implicate that regulation of TA is related to hTERT mRNA isoform expression, and that regulation is different between the cell immortalization and tumorigenesis. (C) 2002 Elsevier Science Ltd.