BACKGROUND: Inhibition of cyclin-dependent kinases 4 (CDK4) activity by Small-interfering RNA (siRNA) has been demonstrated as one of the promising approaches to treat cancer. MATERIALS AND METHODS: CDK4 siRNA was packaged in a lipid nanoparticle (LNP)-based delivery system that consists of an ionizable cationic lipid, helper lipid and polyethylene glycol (PEG)-lipid. The physical properties, including the size and surface charge of LNP-siRNA, were characterized by dynamic light scattering and zeta potential measurements. The biological activities of LNP-siRNA, including the cellular uptake, CDK4 expression and G(1) cell cycle arrest, in both HeLa cervical cancer and MDA-MB-468 breast cancer cells were evaluated by flow cytometry, confocal microscopy, quantitative reverse transcription PCR (qRT-PCR) and Western blotting, respectively. RESULTS: The new LNP-mediated siRNA delivery demonstrated enhanced cellular uptake. Compared with free siRNA and lipofectamine-formulated siRNA, cells treated by LNP-CDK4 siRNA exhibited significant G(1) cell cycle arrest, which was consistent with efficient down-regulation of CDK4 at both mRNA and protein levels. CONCLUSION: Gene silencing of CDK4 by LNP-siRNA offers an alternative strategy for CDK4-based cancer therapy. The new LNP can be used for efficient delivery of siRNA in vitro.
BACKGROUND: Inhibition of cyclin-dependent kinases 4 (CDK4) activity by Small-interfering RNA (siRNA) has been demonstrated as one of the promising approaches to treat cancer. MATERIALS AND METHODS:CDK4 siRNA was packaged in a lipid nanoparticle (LNP)-based delivery system that consists of an ionizable cationic lipid, helper lipid and polyethylene glycol (PEG)-lipid. The physical properties, including the size and surface charge of LNP-siRNA, were characterized by dynamic light scattering and zeta potential measurements. The biological activities of LNP-siRNA, including the cellular uptake, CDK4 expression and G(1) cell cycle arrest, in both HeLa cervical cancer and MDA-MB-468 breast cancer cells were evaluated by flow cytometry, confocal microscopy, quantitative reverse transcription PCR (qRT-PCR) and Western blotting, respectively. RESULTS: The new LNP-mediated siRNA delivery demonstrated enhanced cellular uptake. Compared with free siRNA and lipofectamine-formulated siRNA, cells treated by LNP-CDK4 siRNA exhibited significant G(1) cell cycle arrest, which was consistent with efficient down-regulation of CDK4 at both mRNA and protein levels. CONCLUSION: Gene silencing of CDK4 by LNP-siRNA offers an alternative strategy for CDK4-based cancer therapy. The new LNP can be used for efficient delivery of siRNA in vitro.
Authors: Xinmei Wang; Bo Yu; Wei Ren; Xiaokui Mo; Chenguang Zhou; Hongyan He; Huliang Jia; Lu Wang; Samson T Jacob; Robert J Lee; Kalpana Ghoshal; L James Lee Journal: J Control Release Date: 2013-10-08 Impact factor: 9.776
Authors: Yun Wu; Junyu Ma; Parker S Woods; Nicholas M Chesarino; Chang Liu; L James Lee; Serge P Nana-Sinkam; Ian C Davis Journal: J Control Release Date: 2015-02-14 Impact factor: 9.776
Authors: Xi Zhao; Xiaomeng Huang; Xinmei Wang; Yun Wu; Ann-Kathrin Eisfeld; Sebastian Schwind; Daniel Gallego-Perez; Pouyan E Boukany; Guido I Marcucci; Ly James Lee Journal: Adv Sci (Weinh) Date: 2015-07-16 Impact factor: 16.806
Authors: Shu-Hao Hsu; Bo Yu; Xinmei Wang; Yuanzhi Lu; Carl R Schmidt; Robert J Lee; L James Lee; Samson T Jacob; Kalpana Ghoshal Journal: Nanomedicine Date: 2013-05-30 Impact factor: 5.307