Bioreducible polyethylenimine-delivered siRNA targeting human telomerase reverse transcriptase inhibits HepG2 cell growth in vitro and in vivo.
One new siRNA sequence was found efficient for human telomerase reverse transcriptase (hTERT) gene silencing in vitro in five types of human cancer cells. Then, a biodegradable polyethylenimine containing multiple disulfide bonds (SS-PEI) was successfully applied as a potent non-viral carrier for intracellular delivery of the hTERT siRNA in vitro and in vivo. The SS-PEI could strongly bind siRNA to form nano-sized and positively-charged complexes, but which were readily destabilized to sufficiently release siRNA in a reducing environment. Transfection experiments showed that the complexes of SS-PEI/hTERT siRNA were able to transfect HepG2 cells in vitro, inducing reduced levels of hTERT mRNA and hTERT protein, decreased telomerase activity, cell growth inhibition and significant cell apoptosis. Besides, treatment with the complexes of SS-PEI/hTERT siRNA could inhibit HepG2 tumor growth in a xenograft mouse model. Importantly, the SS-PEI revealed relatively low cytotoxicity in vitro and at an appropriate dose had no adverse effect on liver and kidney functions in vivo. The results of this study indicate that SS-PEI/siRNA-induced hTERT gene silencing provides a promising method for human cancer gene therapy.
Department of Imaging, Tongji Hospital of Tongji University, Shanghai, 200065, PR China.
Wei
W
Xia
Peijun
P
Wang
Chao
C
Lin
Zhuoquan
Z
Li
Xiaolong
X
Gao
Guoliang
G
Wang
Xiaohu
X
Zhao
D000818:Animals; D045744:Cell Line, Tumor; D004337:Drug Carriers / Q000008:administration & dosage* / Q000493:pharmacokinetics; D020868:Gene Silencing; D015316:Genetic Therapy; D006801:Humans; D007668:Kidney / Q000187:drug effects / Q000378:metabolism; D008099:Liver / Q000187:drug effects / Q000378:metabolism; D008297:Male; D051379:Mice; D008819:Mice, Nude; D009369:Neoplasms / Q000188:drug therapy / Q000473:pathology; D011094:Polyethyleneimine / Q000008:administration & dosage* / Q000493:pharmacokinetics; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000493:pharmacokinetics; D051381:Rats; D019098:Telomerase / Q000235:genetics* / Q000378:metabolism; D014018:Tissue Distribution; D047368:Tumor Burden / Q000187:drug effects; D023041:Xenograft Model Antitumor Assays
D004337:Drug Carriers; D034741:RNA, Small Interfering; D011094:Polyethyleneimine; C509186:TERT protein, human; D019098:Telomerase