使用“智能”颗粒沉默上皮癌细胞中的bcl-2表达。

Silencing bcl-2 expression in epithelial cancer cells using "smart" particles.

作者信息

Lin Yen-Ling, Jiang Guohua, Zhang Zhaocheng, Nör Jacques E, ElSayed Mohamed E H

机构信息

Department of Biomedical Engineering, University of Michigan, 1101 Beal Ave., Ann Arbor, MI 48109, USA.

Department of Cariology, Restorative Sciences, and Endodontics, University of Michigan, 1011 N. University Ave., Ann Arbor, MI 48109, USA.

出版信息

J Funct Biomater. 2014 Sep 16;5(3):167-82. doi: 10.3390/jfb5030167.

DOI:10.3390/jfb5030167

PMID:25229941

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4192611/

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.

摘要

靶向抗凋亡Bcl-2蛋白的小干扰RNA（siRNA）被证明可降低其表达并引发癌细胞死亡。我们使用可降解、对pH敏感的梳状[P（EAA-co-BMA）-b-PNASI-g-P（HMA-co-TMAEMA）]聚合物将抗Bcl-2 siRNA浓缩成“智能”颗粒，这些颗粒被证明可将其货物转运穿过内体膜并进入HeLa和UM-SCC-17B癌细胞的细胞质。用抗Bcl-2颗粒处理HeLa和UM-SCC-17B癌细胞，然后分别使用qRT-PCR和蛋白质印迹法对Bcl-2 mRNA和蛋白质水平进行定量。“智能”抗Bcl-2颗粒分别将HeLa细胞中的Bcl-2 mRNA和蛋白质水平选择性抑制50%-60%和79%-81%。同样，“智能”抗Bcl-2颗粒在与UM-SCC-17B癌细胞孵育48、72和96小时后，分别将Bcl-2 mRNA水平抑制30%、40%和20%。在处理72小时后，UM-SCC-17B癌细胞中的Bcl-2蛋白表达被抑制了30%。结果表明，对pH敏感的梳状聚合物复合抗Bcl-2 siRNA形成“智能”纳米颗粒，可将其货物递送至HeLa和UM-SCC-17B癌细胞的细胞质中，在mRNA和蛋白质水平上导致Bcl-2表达降低。