title: RNA interference remarkably suppresses bcl-2 gene expression in cancer cells in vitro and in vivo.
abstract: Bcl-2 is an anti-apoptotic protein. If the level of Bcl-2 protein can be reduced sufficiently in tumors using RNA interference (RNAi) to target the gene message, the apoptosis of tumor cells may be promoted. In this study, we synthesized 19 nucleotides (nts) small interference RNA (siRNA) constructs suppressing bcl-2 gene expression in human tumor cells (HeLaB2 and BGC-823 cell lines) in vitro. The bcl-2 gene expression levels were significantly reduced when these siRNA were transfected into experimental two tumor cells for 72 hours. The apoptosis process was also examined in the tumor cells. Here we synthesized siRNA from a DNA template under the control of the RNA polymerase III promoter in transfected tumor cells. Using this DNA vector-based approach, we found that the siRNA efficiently and specifically inhibited the synthesis of protein encoded by the bcl-2 gene in HeLaB2 and BGC-823 tumor cells. Tumor growth was inhibited by 66.5% with 2mg/kg pSilencer 3.1H1-bcl-2 in mouse liver tumor-bearing BALB/c mice. This approach may prove to be a valuable clinical technique for the analysis of specific gene functions and gene therapy of malignant tumors that utilize the bcl-2 gene via RNA interference.
authors.affiliation: Medical School and State Key Loboratory of Pharmaceutical Biotechnology, Nanjing University, China.
authors.forename: Geng-Feng
authors.initials: GF
authors.lastname: Fu
authors.forename: Xue-Hong
authors.initials: XH
authors.lastname: Lin
authors.forename: Qing-Wang
authors.initials: QW
authors.lastname: Han
authors.forename: Yan-Rong
authors.initials: YR
authors.lastname: Fan
authors.forename: Ye-Fen
authors.initials: YF
authors.lastname: Xu
authors.forename: Dan
authors.initials: D
authors.lastname: Guo
authors.forename: Gen-Xing
authors.initials: GX
authors.lastname: Xu
authors.forename: Ya-Yi
authors.initials: YY
authors.lastname: Hou
mesh_terms: D000818:Animals; D017209:Apoptosis; D045744:Cell Line, Tumor; D015536:Down-Regulation; D015870:Gene Expression; D019254:Genes, bcl-2 / Q000235:genetics*; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D009369:Neoplasms / Q000737:chemistry / Q000235:genetics / Q000628:therapy*; D010957:Plasmids / Q000235:genetics; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000032:analysis / Q000037:antagonists & inhibitors* / Q000235:genetics; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D037941:Silencer Elements, Transcriptional / Q000235:genetics; D023041:Xenograft Model Antitumor Assays
chemical_list: D019253:Proto-Oncogene Proteins c-bcl-2; D034741:RNA, Small Interfering