Basic Research
Copyright ©2005 Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. May 21, 2005; 11(19): 2912-2915
Published online May 21, 2005. doi: 10.3748/wjg.v11.i19.2912
Effect of vector-expressed shRNAs on hTERT expression
Ying Guo, Jun Liu, Ying-Hui Li, Tian-Bao Song, Jing Wu, Cai-Xia Zheng, Cai-Fang Xue
Ying Guo, Jing Wu, Cai-Xia Zheng, Department of Obstetrics and Gynecology, Second Hospital of Xi’an Jiaotong University, Xi’an 710001, Shaanxi Province, China
Jun Liu, Ying-Hui Li, Cai-Fang Xue, Department of Etiology, Fourth Military Medical University, Xi’an 710033, Shaanxi Province, China
Tian-Bao Song, Department of Histology and Embryology, Medical School of Xi’an Jiaotong University, Xi’an 710061, Shaanxi Province, China
Author contributions: All authors contributed equally to the work.
Supported by the Shaanxi Province Natural Science Foundation, No. 2003C217
Correspondence to: Dr. Jun Liu, Department of Etiology, Fourth Military Medical University, Xi’an 710033, Shaanxi Province, China. etiology@fmmu.edu.cn
Telephone: +86-29-83374536 Fax: +86-29-83374594
Received: June 29, 2004
Revised: June 30, 2004
Accepted: July 22, 2004
Published online: May 21, 2005
Abstract

AIM: To study the effect of short hairpin RNAs (shRNAs) expressed from DNA vector on hTERT expression.

METHODS: Oligonucleotides coding for four shRNAs against hTERT were cloned into a mammalian shRNA expression vector pUC18U6 to form pUC18U6ht1-4, which were then introduced into HepG2 cells by using liposome-mediated transfection. HepG2 cells transfected by pUC18U6 and pUC18U6GFPsir, which expressed shRNA against green fluorescent protein (GFP), were used as controls. hTERT mRNA in the transfected cells were quantified by using real-time fluorescent RT-PCR.

RESULTS: Among the four shRNAs against hTERT, two decreased the hTERT mRNA level. Compared with the controls, pUC18U6ht which expressed the two shRNAs reduced hTERT mRNA by 39% and 49% (P<0.05).

CONCLUSION: hTERT expression is inhibited by the shRNAs expressed from the DNA vector.

Keywords: RNA interference; Short hairpin RNA; Telomerase