Effect of vector-expressed shRNAs on hTERT expression.
OBJECTIVE
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).


CONCLUSIONS
hTERT expression is inhibited by the shRNAs expressed from the DNA vector.
Guo
Ying
Y
Department of Etiology, Fourth Military Medical University, Xi'an 710033, Shaanxi Province, China.
Liu
Jun
J
Li
Ying-Hui
YH
Song
Tian-Bao
TB
Wu
Jing
J
Zheng
Cai-Xia
CX
Xue
Cai-Fang
CF
D045744:Cell Line, Tumor; D004268:DNA-Binding Proteins; D005786:Gene Expression Regulation; D015316:Genetic Therapy / Q000379:methods*; D018197:Hepatoblastoma / Q000628:therapy*; D006801:Humans; D008113:Liver Neoplasms / Q000628:therapy*; D010957:Plasmids / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics*; D019098:Telomerase / Q000235:genetics*; D014162:Transfection
D004268:DNA-Binding Proteins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D019098:Telomerase