title: Silencing bcl-2 expression in epithelial cancer cells using "smart" particles.
abstract: Short interfering RNA (siRNA) targeted against anti-apoptotic Bcl-2 protein proved to knockdown its expression and trigger cancer cell death. We used degradable, pH-sensitive, comb-like [P(EAA-co-BMA)-b-PNASI-g-P(HMA-co-TMAEMA)] polymer to condense anti-Bcl-2 siRNA into "smart" particles, which proved to shuttle their cargo past the endosomal membrane and into the cytoplasm of HeLa and UM-SCC-17B cancer cells. HeLa and UM-SCC-17B cancer cells were treated with anti-Bcl-2 particles followed by quantifying Bcl-2 mRNA and protein levels using qRT-PCR and western blotting, respectively. "Smart" anti-Bcl-2 particles selectively suppress Bcl-2 mRNA and protein levels in HeLa cells by 50%-60% and 79%-81%, respectively. Similarly, "smart" anti-Bcl-2 particles inhibited Bcl-2 mRNA levels by 30%, 40%, and 20% upon incubation with UM-SCC-17B cancer cells for 48, 72, and 96 h, respectively. Bcl-2 protein expression in UM-SCC-17B cancer cells was inhibited by 30% after treatment for 72 h. Results show that pH-sensitive comb-like polymer complex anti-Bcl-2 siRNA forming "smart" nanoparticles that deliver their cargo into the cytoplasm of HeLa and UM-SCC-17B cancer cells causing Bcl-2 knockdown at the mRNA and protein levels.
authors.affiliation: Department of Biomedical Engineering, University of Michigan, 1101 Beal Ave., Ann Arbor, MI 48109, USA. yenling225@gmail.com.
authors.forename: Yen-Ling
authors.initials: YL
authors.lastname: Lin
authors.affiliation: Department of Biomedical Engineering, University of Michigan, 1101 Beal Ave., Ann Arbor, MI 48109, USA. yenling225@gmail.com.
authors.forename: Guohua
authors.initials: G
authors.lastname: Jiang
authors.affiliation: Department of Cariology, Restorative Sciences, and Endodontics, University of Michigan, 1011 N. University Ave., Ann Arbor, MI 48109, USA. zczhang@umich.edu.
authors.forename: Zhaocheng
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: Department of Cariology, Restorative Sciences, and Endodontics, University of Michigan, 1011 N. University Ave., Ann Arbor, MI 48109, USA. jenor@umich.edu.
authors.forename: Jacques E
authors.initials: JE
authors.lastname: Nör
authors.affiliation: Department of Biomedical Engineering, University of Michigan, 1101 Beal Ave., Ann Arbor, MI 48109, USA. yenling225@gmail.com.
authors.forename: Mohamed E H
authors.initials: ME
authors.lastname: ElSayed