Oligomer-mediated modulation of hTERT alternative splicing induces telomerase inhibition and cell growth decline in human prostate cancer cells
Oligomer-mediated modulation of hTERT alternative splicing induces telomerase inhibition and cell growth decline in human prostate cancer cells
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DOI：
10.1007/s00018-004-4062-7
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发表时间：
2004-07-01
影响因子：
8
通讯作者：
Zaffaroni, N
中科院分区：
文献类型：
--
作者：
Brambilla, C;Folini, M;Zaffaroni, N
人类细胞中端粒酶的表达受到端粒酶逆转录酶（hTERT）转录和选择性剪接等多种机制的严格控制。在这项研究中，我们证明了通过使用2'- o -甲基rna磷酸化寡核苷酸靶向hTERT前mrna中位于内含子5和外显子6之间的剪接位点来调节DU145人前列腺癌细胞中hTERT剪接模式的可能性。寡核苷酸暴露18 h诱导全长hTERT转录物减少，同时选择性剪接转录物增加，导致端粒酶催化活性显著抑制。此外，暴露于R7寡聚物（诱导最明显的hTERT剪接模式调节和最大的端粒酶抑制）导致DU145细胞生长显著减少，并在治疗后2天开始诱导凋亡。这些数据支持了hTERT表达下调可以对肿瘤细胞生长产生短期影响的概念，这种影响与端粒缩短无关。
The expression of telomerase in human cells is strictly controlled by multiple mechanisms including transcription and alternative splicing of telomerase reverse transcriptase (hTERT). In this study, we demonstrated the possibility of modulating the hTERT splicing pattern in DU145 human prostate carcinoma cells through the use of 2'-O-methyl-RNA phosphorothioate oligonucleotides targeting the splicing site located between intron 5 and exon 6 in the hTERT pre-mRNA. An 18-h oligonucleotide exposure induced a decrease in the full-length hTERT transcript and a concomitant increase in the alternatively spliced transcripts, which resulted in significant inhibition of telomerase catalytic activity. Moreover, exposure to the R7 oligomer (which induced the most pronounced modulation of the hTERT splicing pattern and the greatest telomerase inhibition) caused a marked reduction in DU145 cell growth and the induction of apoptosis starting 2 days after treatment. Such data support the concept that down-regulation of hTERT expression can cause short-term effects on tumour cell growth, which are telomere-shortening independent.