title: Guanidinylated 3-gluconamidopropyl methacrylamide-s-3-aminopropyl methacrylamide copolymer as siRNA carriers for inhibiting human telomerase reverse transcriptase expression.
abstract: In this report, a series of well-defined glucose- and guanidine-based cationic copolymers as gene carriers were developed to inhibit human telomerase reverse transcriptase (hTERT) gene expression. First of all, guandinylated 3-gluconamidopropyl methacrylamide-s-3-aminopropyl methacrylamide copolymers (guanidinylated GAPMA-s-APMA, abbreviated as GGA) were prepared via aqueous reversible addition--fragmentation chain transfer polymerization (RAFT). Then, three target hTERT siRNA TERT-1, TERT-2 and TERT-3 were designed and combined with GGA copolymers to form siRNA/GGA polyplexes. The polyplexes were examined by dynamic light scattering and agarose gel electrophoresis. The results indicated that GGA copolymers can condense siRNA effectively to form particles with the diameter from 157 nm to 411 nm and zeta potential values in the range from +3.7 to +15.8 mV at various charge ratios (N/P). The MTT assay data of siRNA/GGA polyplexes on human hepatocellular liver carcinoma cells (HepG2) indicated that GGA copolymer had better cell viabilities than polyethylenimine (PEI). Furthermore, the transfection of siRNA/GGA polyplexes was detected by real-time quantitative PCR (RT-qPCR) in HepG2. It was found that siRNA/GGA polyplexes could effectively silence hTERT mRNA expression in serum-free media (p<0.01). In the presence of serum, the hTERT mRNA expression in HepG2 cells have significant difference (p<0.01) between siRNA/GGA3 polyplexes and blank. The results showed that the GAPMA component can reduce the aggregation of protein in serum media. Therefore, the enhancement of transfection may be attributed to the combination of guadino groups and glucose component. And, the guandinylated 3-gluconamidopropyl methacrylamide-s-3-aminopropyl methacrylamide copolymers might be promise in gene delivery.
authors.affiliation: Biomaterials and Drug Delivery Laboratories, School of Chemistry and Chemical Engineering, Southeast University , Nanjing , China and.
authors.forename: Yang
authors.initials: Y
authors.lastname: Wu
authors.forename: Jinkai
authors.initials: J
authors.lastname: Ji
authors.forename: Ran
authors.initials: R
authors.lastname: Yang
authors.forename: Xiaoqiang
authors.initials: X
authors.lastname: Zhang
authors.forename: Yuanhui
authors.initials: Y
authors.lastname: Li
authors.forename: Yuepu
authors.initials: Y
authors.lastname: Pu
authors.forename: Xinsong
authors.initials: X
authors.lastname: Li
mesh_terms: D000178:Acrylamides / Q000737:chemistry; D006528:Carcinoma, Hepatocellular / Q000235:genetics; D002470:Cell Survival; D004587:Electrophoresis, Agar Gel; D015971:Gene Expression Regulation, Enzymologic; D020868:Gene Silencing; D018014:Gene Transfer Techniques; D005947:Glucose / Q000737:chemistry; D019791:Guanidine / Q000737:chemistry; D056945:Hep G2 Cells; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics; D010316:Particle Size; D011108:Polymers / Q000138:chemical synthesis / Q000737:chemistry*; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000008:administration & dosage*; D060888:Real-Time Polymerase Chain Reaction; D019098:Telomerase / Q000235:genetics*; D014162:Transfection
chemical_list: D000178:Acrylamides; D011108:Polymers; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D019098:Telomerase; D005947:Glucose; D019791:Guanidine; C045985:methacrylamide