Human normal T lymphocytes and lymphoid cell lines do express alternative splicing variants of human telomerase reverse transcriptase (hTERT) mRNA

Jalink, Marit; Ge, Zheng; Liu, Cheng; Björkholm, Magnus; Gruber, Astrid; Xu, Dawei

doi:10.1016/j.bbrc.2006.12.149

Cited by 22 publications

(25 citation statements)

References 24 publications

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“…Alternatively spliced hTERT mRNA contributes to telomerase regulation under certain settings, and it has been established that the full-length hTERT transcripts, rather than any other variants, is translated into a functional protein required for active telomerase (5,6,(24)(25)(26)(27)(28)(29)(30). The recent study has indicated that hypoxia drives preferential expression of the full-length hTERT mRNA in ovary cancer cells (10).…”

Section: Discussionmentioning

confidence: 99%

“…It is known that the alternative splicing of hTERT transcription participates in controlling telomerase activity (5,(24)(25)(26)(27)(28)(29)(30)(31), and that a recent study suggests a transcriptional switch to the full-length hTERT mRNA in ovary cancer cells in response to hypoxia (10). Therefore, we asked whether this was the case in RCC cells by determining the expression profile of hTERT mRNA variants.…”

Section: Hypoxic Treatment Up-regulates Overall Htert Mrna Expressionmentioning

confidence: 99%

“…The PCR primer sequences specific for all the variants of hTERT mRNA (accession no. AF015950) were 5 ¶-CGGAAGAGTGTCTGGAGCAA-3 ¶ and 5 ¶-GGATGAA-GCGGAGTCTGGA-3 ¶ (24,29), and the resultant amplicon was 145 bp long. The amplification of the alternatively spliced hTERT mRNA was done with the primers 2164S (5 ¶-GCCTGAGCTGTACTTTGTCAA-3 ¶) and 2620AS (5 ¶-CGCAAACAGCTTGTTCTCCATGTC-3 ¶; refs.…”

Section: Rna Extraction and Rt-pcrmentioning

confidence: 99%

“…The amplification of the alternatively spliced hTERT mRNA was done with the primers 2164S (5 ¶-GCCTGAGCTGTACTTTGTCAA-3 ¶) and 2620AS (5 ¶-CGCAAACAGCTTGTTCTCCATGTC-3 ¶; refs. 24,25,29). This single primer set allowed detection of all differently spliced variants, including the full-length hTERT transcript simultaneously (25).…”

Section: Rna Extraction and Rt-pcrmentioning

confidence: 99%

“…The cells with different treatments were harvested, and total cellular proteins were extracted with SDS lysis buffer (29). About 30 Ag of the proteins were resolved in SDS-PAGE and transferred to an intracellulose membrane.…”

Section: Western Blotmentioning

confidence: 99%

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The Opposing Effect of Hypoxia-Inducible Factor-2α on Expression of Telomerase Reverse Transcriptase

Lou¹,

Chen

Jalink

et al. 2007

Molecular Cancer Research

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Hypoxia-inducible factor-1A (HIF-1A) has been implicated in the transcriptional regulation of the telomerase reverse transcriptase (hTERT) gene expression and telomerase activity, essential elements for cellular immortalization and transformation. However, controversial results were obtained in different studies. Moreover, it is totally unclear whether HIF-2A, the paralog of HIF-1A, plays a role in regulating hTERT expression. In the present study, we found that hypoxic treatment enhanced hTERT mRNA expression and telomerase activity in three renal cell carcinoma (RCC) cell lines with different genetic backgrounds. Both HIF-1A and HIF-2A were capable of significantly increasing the hTERT promoter activity in these cells. Moreover, depleting HIF-2A led to a down-regulation of hTERT mRNA level in RCC A498 cells expressing constitutive HIF-2A. It was found that HIF-2A bound to the hTERT proximal promoter and enhanced the recruitment of the histone acetyltransferase p300 and histone H3 acetylation locally in A498 cells treated with hypoxia. Increased levels of hTERT mRNA were observed in two of three hypoxia-treated malignant glioma cell lines. However, HIF-1A stimulated whereas HIF-2A inhibited the hTERT promoter activity in these glioma cell lines. Ectopic expression of HIF-2A resulted in diminished hTERT expression in glioma cells. Collectively, HIF-1A activates hTERT and telomerase expression in both RCC and glioma cells, and HIF-2A enhances hTERT expression in RCC cells, whereas it represses the hTERT transcription in glioma cells.

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