title: Gene therapy for colon cancer by adeno-associated viral vector-mediated transfer of survivin Cys84Ala mutant.
abstract: OBJECTIVE
  Reactivation of survivin expression is involved in carcinogenesis and angiogenesis in colon cancer. Previous in vitro studies showed that mutation of the cysteine residue at position 84 (Cys84Ala) of survivin generates a dominant-negative mutant that triggers mitotic catastrophe and apoptosis. We investigated the therapeutic effect of the adeno-associated virus (AAV)-mediated survivin mutant (Cys84Ala) on colon cancer.
  
  
  METHODS
  Survivin mutant (Cys84Ala) (Sur-Mut(Cys84Ala)) was cloned into the AAV expression vector pAM/CAG-WPRE.poly(A) to generate recombinant AAV-Sur-Mut(Cys84Ala) virus. Cell proliferation, apoptosis, mitotic catastrophe, and tumor growth were measured in vitro and in vivo.
  
  
  RESULTS
  Transduction of colon cancer cells with rAAV-Sur-Mut(Cys84Ala) inhibited cell proliferation and induced apoptosis and mitotic catastrophe in vitro. rAAV-Sur-Mut(Cys84Ala) sensitized colon cancer cells to chemotherapeutic drugs. Furthermore, expression of survivin mutant mediated by AAV inhibited tumorigenesis in colon cancer cells. Intratumoral injection of rAAV-Sur-Mut(Cys84Ala) significantly induced apoptosis and mitotic catastrophe and inhibited angiogenesis and tumor growth in a colon cancer xenograft model in vivo. No obvious cytotoxicity to other tissues was observed. More importantly, rAAV-Sur-Mut(Cys84Ala) expression strongly enhanced the antitumor activity of 5-Fluorouracil (5-FU), resulting in regression of established tumors.
  
  
  CONCLUSIONS
  Our results showed that rAAV-Sur-Mut(Cys84Ala) induced apoptosis and mitotic catastrophe and inhibited tumor angiogenesis and tumor growth. Thus, use of AAV-mediated survivin mutant (Cys84Ala) is a promising strategy in colon cancer gene therapy.
authors.affiliation: Department of Gastroenterology, Rui-jin Hospital, Shanghai, P.R. China.
authors.forename: Shui Ping
authors.initials: SP
authors.lastname: Tu
authors.forename: Jian Tao
authors.initials: JT
authors.lastname: Cui
authors.forename: Peter
authors.initials: P
authors.lastname: Liston
authors.forename: Xiao
authors.initials: X
authors.lastname: Huajiang
authors.forename: Ruian
authors.initials: R
authors.lastname: Xu
authors.forename: Marie C M
authors.initials: MC
authors.lastname: Lin
authors.forename: Yan Bo
authors.initials: YB
authors.lastname: Zhu
authors.forename: Bing
authors.initials: B
authors.lastname: Zou
authors.forename: Samuel S M
authors.initials: SS
authors.lastname: Ng
authors.forename: Shi Hu
authors.initials: SH
authors.lastname: Jiang
authors.forename: Harry H X
authors.initials: HH
authors.lastname: Xia
authors.forename: Wai Man
authors.initials: WM
authors.lastname: Wong
authors.forename: Annie O O
authors.initials: AO
authors.lastname: Chan
authors.forename: Man Fung
authors.initials: MF
authors.lastname: Yuen
authors.forename: Shiu Kum
authors.initials: SK
authors.lastname: Lam
authors.forename: Hsiang Fu
authors.initials: HF
authors.lastname: Kung
authors.forename: Benjamin C Y
authors.initials: BC
authors.lastname: Wong
mesh_terms: D000409:Alanine; D019943:Amino Acid Substitution; D017209:Apoptosis; D002455:Cell Division; D045744:Cell Line, Tumor; D003110:Colonic Neoplasms / Q000235:genetics* / Q000473:pathology / Q000628:therapy*; D003545:Cysteine; D000229:Dependovirus / Q000235:genetics*; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins / Q000235:genetics*; D008938:Mitosis; D020125:Mutation, Missense; D009363:Neoplasm Proteins; D000077022:Survivin
chemical_list: C107489:BIRC5 protein, human; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins; D009363:Neoplasm Proteins; D000077022:Survivin; D003545:Cysteine; D000409:Alanine