中华医学杂志（英文版）2002，Vol.115Issue(8)：1196-1200,5.
稳定表达反义hTR抑制人胰腺癌细胞生长
Stable expression of antisense hTR inhibits in vitro pancreatic cancer cell growth
滕理送 1TJFahey1
作者信息
- 1. 浙江大学医学院附属第一医院,杭州,310003
- 折叠
摘要
Abstract
Objective To clarify growth inhibition in pancreatic cancer cells by interference with the hTR component of the telomerase reverse transcriptase enzymatic complex. Methods A 593 bp full length hTR cDNA was subcloned into a mammalian expression vector pcDNA3.1(-) in the antisense orientation to construct an antisense hTR expression plasmid. These were introduced into panc1 cells, a human pancreatic carcinoma cell line, by lipofectin and G418-resistant stable transformants were expanded. Resulting stable clones were screened for the presence of the hTR insert by PCR with T7 and BGH reverse primers located on the flanks of the multiclonal site of the pcDNA3.1 vector. Cell growth rate, hTR expression, telomerase activity and anchorage-independent growth properties were analyzed. Results Significant downregulation of endogenous hTR was evident in the antisense-hTR transformed cells and telomerase activity was markedly decreased compared to control cells in standard TRAP assays. Furthermore, cell proliferation and the anchorage-independent growth ability in antisense-hTR expressing cells were significantly decreased compared with control parental cells. However, no crisis or senescence phenomena were observed. Conclusions These data indicate that hTR may be a critical component of human telomerase activity and suggest that downregulation of the RNA component of human telomerase is a possible target for anticancer strategies.关键词
端粒酶 人端粒酶模板RNA 反义 胰腺癌细胞系Key words
telomerase/hTR/antisense/pancreatic carcinoma cell line分类
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滕理送,TJFahey..稳定表达反义hTR抑制人胰腺癌细胞生长[J].中华医学杂志（英文版）,2002,115(8):1196-1200,5.
