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Quantification of Alternative Splicing Variants of Human Telomerase Reverse Transcriptase and Correlations with Telomerase Activity in Lung Cancer
Liu, Yan; Wu, Bing-quan; Zhong, Hao-hao; Tian, Xin-xia; Fang, Wei-gang
刊名PLOS ONE
2012-06-18
DOI10.1371/journal.pone.0038868
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
WOS记录号WOS:000305583300062
项目编号2010CB529402
资助机构Ministry of Science and Technology of China
引用统计
被引频次:17[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.bjmu.edu.cn/handle/400002259/68262
专题北京大学基础医学院
北京大学医学部管理机构_主任办公室党委办公室
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
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GB/T 7714
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).
APA Liu, Yan,Wu, Bing-quan,Zhong, Hao-hao,Tian, Xin-xia,&Fang, Wei-gang.(2012).Quantification of Alternative Splicing Variants of Human Telomerase Reverse Transcriptase and Correlations with Telomerase Activity in Lung Cancer.PLOS ONE,7(6).
MLA Liu, Yan,et al."Quantification of Alternative Splicing Variants of Human Telomerase Reverse Transcriptase and Correlations with Telomerase Activity in Lung Cancer".PLOS ONE 7.6(2012).
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