title: A novel colon cancer gene therapy using rAAV‑mediated expression of human shRNA-FHL2.
abstract: FHL2 (Four and a half LIM-only protein 2) has been identified as an oncogene in colon cancer and suppression of FHL2 induces cell differentiation and tumorigenesis in colon cancer cell lines. The aim of this study was to develop a novel and effective approach to knockdown FHL2, which can serve as a promising target of colon cancer therapy. Recombinant adeno-associated virus (rAAV) was generated bearing with FHL2-shRNA and transfected into LoVo cells. Cell cycle and growth were assessed. The interaction between FHL2 and G0/G1 cell cycle and growth was evaluated by flow cytometry, western blot analysis and WST-1 assay. We showed that suppression of FHL2 by rAAV-shRNA induced G0/G1 cell cycle arrest and inhibited cell growth. Apoptosis-related proteins and their activity was investigated at the same time. rAAV-FHL2‑shRNA activated intrinsic and extrinsic apoptotic pathways and increased cell susceptibility to apoptotic stimuli by 5-FU. Moreover, a xenograft model was established to explore rAAV-FHL2-shRNA with 5-FU mediated tumorigenesis in vivo. A strong anti-tumorigenic effect of rAAV-FHL2-shRNA was shown in nude mice and this antitumor effect was enhanced when combined with 5-FU treatment. These findings implicate FHL2 as a cell cycle and growth modulator and thus inhibit apoptosis in colon cancer cells. rAAV-shRNA-FHL2 may serve as a novel and potent therapeutic or 5-FU co-therapeutic agent for colon cancer.
authors.affiliation: Guangdong Provincial Key Laboratory of Gastroenterology, Department of Digestive Medicine, Nanfang Hospital, Southern Medical University, Guangzhou 510515, P.R. China.
authors.forename: Yao
authors.initials: Y
authors.lastname: Wu
authors.forename: Zheng
authors.initials: Z
authors.lastname: Guo
authors.forename: Di
authors.initials: D
authors.lastname: Zhang
authors.forename: Wenjing
authors.initials: W
authors.lastname: Zhang
authors.forename: Qingqing
authors.initials: Q
authors.lastname: Yan
authors.forename: Xinpeng
authors.initials: X
authors.lastname: Shi
authors.forename: Mengnan
authors.initials: M
authors.lastname: Zhang
authors.forename: Yingying
authors.initials: Y
authors.lastname: Zhao
authors.forename: Yali
authors.initials: Y
authors.lastname: Zhang
authors.forename: Bo
authors.initials: B
authors.lastname: Jiang
authors.forename: Tianming
authors.initials: T
authors.lastname: Cheng
authors.forename: Yang
authors.initials: Y
authors.lastname: Bai
authors.forename: Jide
authors.initials: J
authors.lastname: Wang
mesh_terms: D000818:Animals; D017209:Apoptosis; D015153:Blotting, Western; D002453:Cell Cycle; D049109:Cell Proliferation; D003110:Colonic Neoplasms / Q000235:genetics / Q000473:pathology / Q000628:therapy*; D000229:Dependovirus / Q000235:genetics*; D005260:Female; D005434:Flow Cytometry; D015316:Genetic Therapy; D006801:Humans; D060850:LIM-Homeodomain Proteins / Q000037:antagonists & inhibitors / Q000235:genetics*; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009124:Muscle Proteins / Q000037:antagonists & inhibitors / Q000235:genetics*; D034741:RNA, Small Interfering / Q000235:genetics*; D014157:Transcription Factors / Q000037:antagonists & inhibitors / Q000235:genetics*
chemical_list: C105987:FHL2 protein, human; D060850:LIM-Homeodomain Proteins; D009124:Muscle Proteins; D034741:RNA, Small Interfering; D014157:Transcription Factors