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学科主题: 基础医学
题名:
Quantification of Alternative Splicing Variants of Human Telomerase Reverse Transcriptase and Correlations with Telomerase Activity in Lung Cancer
作者: Liu, Yan; Wu, Bing-quan; Zhong, Hao-hao; Tian, Xin-xia; Fang, Wei-gang
刊名: PLOS ONE
发表日期: 2012-06-18
DOI: 10.1371/journal.pone.0038868
卷: 7, 期:6
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology
类目[WOS]: Multidisciplinary Sciences
研究领域[WOS]: Science & Technology - Other Topics
关键词[WOS]: REPEAT AMPLIFICATION PROTOCOL ; MESSENGER-RNA ; QUANTITATIVE-ANALYSIS ; HTERT EXPRESSION ; DELETION VARIANT ; ASSAY ; GENE ; CARCINOMA ; CELLS ; DIFFERENTIATION
英文摘要:

Telomerase plays important roles in the development and progression of malignant tumors, and its activity is primarily determined by transcriptional regulation of human telomerase reverse transcriptase (hTERT). Several mRNA alternative splicing variants (ASVs) for hTERT have been identified, but it remains unclear whether telomerase activity is directly associated with hTERT splicing transcripts. In this study, we developed novel real-time PCR protocols using molecular beacons and applied to lung carcinoma cell lines and cancerous tissues for quantification of telomerase activity and three essential hTERT deletion transcripts respectively. The results showed that lung carcinoma cell lines consistently demonstrated telomerase activity (14.22-31.43 TPG units per 100 cells) and various hTERT alternative splicing transcripts. For 165 lung cancer cases, telomerase activity showed significant correlation with tumor differentiation (poorly-> moderately-> well-differentiated, P < 0.01) and with histotypes (combined small cell and squamous cell carcinoma > squamous cell carcinoma > adenosquamous carcinoma > adenocarcinoma, P < 0.05). Although the overall hTERT transcripts were detected in all the samples, they were not associated with telomerase activity (r = 0.092, P = 0.24). Telomerase activity was significantly correlated with the transcriptional constituent ratio of alpha-deletion (r = -0.267, P = 0.026), beta-deletion (r = -0.693, P = 0.0001) and gamma-deletion (r = -0.614, P = 0.001). The positive rate and average constituent ratio of beta-deletion transcripts (92.12%, 0.23) were higher than those of alpha-deletion (41.82%, 0.12) or gamma-deletion (16.36%, 0.18) transcripts. The combined small-cell and squamous cell carcinomas expressed less deletion transcripts, especially beta-deletion, than other histotypes, which might explain their higher telomerase activity. In conclusion, the molecular beacon-based real-time PCR protocols are rapid, sensitive and specific methods to quantify telomerase activity and hTERT ASVs. Telomerase activity may serve as a reliable and effective molecular marker to assist the evaluation of histological subtype and differentiation of lung carcinomas. Further studies on hTERT deletion splicing transcripts, rather than the overall hTERT transcripts, may improve our understanding of telomerase regulation.

语种: 英语
所属项目编号: 2010CB529402
项目资助者: Ministry of Science and Technology of China
WOS记录号: WOS:000305583300062
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/68262
Appears in Collections:基础医学院_期刊论文

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作者单位: Peking Univ, Key Lab Carcinogenesis & Translat Res, Sch Basic Med Sci, Minist Educ,Hlth Sci Ctr, Beijing 100871, Peoples R China

Recommended Citation:
Liu, Yan,Wu, Bing-quan,Zhong, Hao-hao,et al. Quantification of Alternative Splicing Variants of Human Telomerase Reverse Transcriptase and Correlations with Telomerase Activity in Lung Cancer[J]. PLOS ONE,2012,7(6).
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