Akt1 silencing efficiencies in lung cancer cells by sh/si/ssiRNA transfection using a reductable polyspermine carrier.
Efforts directed in ameliorating silencing studies with shRNA, siRNA and ssiRNA (siRNA with sticky overhangs) are faltered mainly due to the lack of efficient carrier system. In the present study, we developed reductable polyspermine (RPS) carrier composed of multiple spermine units with disulfide linkages for gene expression and silencing studies. In gene expression studies, EGFP expression was found to be almost 4 folds higher and 20 folds safer with RPS carrier than with PEI25K. Moreover, on systemic administration, RPS exhibited significantly high EGFP expression in mice lungs. Similarly in gene silencing studies, EGFP silencing achieved was nearly 1.5 times superior with RPS carrier than PEI25K. Also, RPS delivered Akt1 shRNA (shAkt), siRNA (siAkt) and ssiRNA (ssiAkt) efficiently silenced oncoprotein Akt1 and thereby decreased A549 cell survival. The degrees of cell survival, proliferation and metastasis were differed with the nature of siRNA treatment. Further study at different time intervals revealed that ssiAkt treatment, although superior to sh/siAkt, was highly transient while, shAkt treatment was uniform and prolong. These finding demonstrate the potential use of RPS carrier in gene expression and silencing studies, and significance of the nature of siRNA employed in cancer study.
Jere
Dhananjay
D
Department of Agricultural Biotechnology, Seoul National University, Seoul, Republic of Korea.
Kim
Ji-Eun
JE
Arote
Rohidas
R
Jiang
Hu-Lin
HL
Kim
You-Kyoung
YK
Choi
Yun Jaie
YJ
Yun
Cheol-Heui
CH
Cho
Myung-Haing
MH
Cho
Chong-Su
CS
D000818:Animals; D017209:Apoptosis; D001692:Biological Transport; D002452:Cell Count; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation; D002470:Cell Survival; D004247:DNA / Q000378:metabolism; D004337:Drug Carriers; D020868:Gene Silencing; D049452:Green Fluorescent Proteins / Q000378:metabolism; D042541:Intracellular Space / Q000378:metabolism; D008175:Lung Neoplasms / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D009682:Magnetic Resonance Spectroscopy; D051379:Mice; D010316:Particle Size; D010957:Plasmids / Q000378:metabolism; D011108:Polymers / Q000138:chemical synthesis / Q000378:metabolism*; D051057:Proto-Oncogene Proteins c-akt / Q000235:genetics* / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism*; D013096:Spermine / Q000138:chemical synthesis / Q000378:metabolism*; D014162:Transfection
D004337:Drug Carriers; D011108:Polymers; D034741:RNA, Small Interfering; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D013096:Spermine; D004247:DNA; D051057:Proto-Oncogene Proteins c-akt