title: In vitro and in vivo effect of 5-FC combined gene therapy with TNF-α and CD suicide gene on human laryngeal carcinoma cell line Hep-2.
abstract: This study was aimed to investigate the effect of combined cancer gene therapy with exogenous tumor necrosis factor-alpha (TNF-α) and cytosine deaminase (CD) suicide gene on laryngeal carcinoma cell line Hep-2 in vitro and in vivo. Transfection of the recombinant eukaryotic vectors of pcDNA3.1 (+) containing TNF-α and/or CD into Hep-2 cells resulted in expression of TNF-α and/or CD gene in vitro. The significant increase in apoptotic Hep-2 cells and decrease of Hep-2 cell proliferation were observed using 5-FC treatment combined with TNF-a expression by CD/5-FC suicide system. Moreover, bystander effect was also observed in the TNF-α and CD gene co-expression group. Laryngeal squamous cell carcinoma (LSCC) mice model was established by using BALB/c mice which different transfected Hep-2 cells with pcDNA3.1 (+) containing TNF-α and/or CD were applied subcutaneously. So these mice are divided into four groups, namely, (1)Hep-2/TIC group; (2)Hep-2/CD group; (3)Hep-2/TNF-α group; (4)Hep-2/0 group. At day 29 after cell inoculation, volume of grafted tumor had significant difference between each two of them (P<0.05). These results showed that the products of combined CD and TNF-α genes inhibited the growth of transplanted LSCC in mice model. So by our observed parameters and many others results, we hypothesized that 5-FC combined gene therapy with TNF-αand CD suicide gene should be an effective treatment on Laryngeal carcinoma.
authors.affiliation: Department of Otolaryngology, First Affiliated Hospital and Otolaryngology Institute, Sun Yat-Sen University, Guangzhou, Guangdong, PR China. clp58612@163.com
authors.forename: Li-Ping
authors.initials: LP
authors.lastname: Chai
authors.forename: Zhang-Feng
authors.initials: ZF
authors.lastname: Wang
authors.forename: Wei-Ying
authors.initials: WY
authors.lastname: Liang
authors.forename: Lei
authors.initials: L
authors.lastname: Chen
authors.forename: Dan
authors.initials: D
authors.lastname: Chen
authors.forename: An-Xun
authors.initials: AX
authors.lastname: Wang
authors.forename: Zhao-Qiang
authors.initials: ZQ
authors.lastname: Zhang
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D002294:Carcinoma, Squamous Cell / Q000188:drug therapy / Q000235:genetics / Q000628:therapy*; D045744:Cell Line, Tumor; D003001:Cloning, Molecular; D043525:Cytosine Deaminase / Q000235:genetics* / Q000378:metabolism; D017931:DNA Primers / Q000235:genetics; D004926:Escherichia coli; D005437:Flucytosine / Q000627:therapeutic use*; D041722:Genes, Transgenic, Suicide / Q000235:genetics*; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006801:Humans; D007822:Laryngeal Neoplasms / Q000188:drug therapy / Q000235:genetics / Q000628:therapy*; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics* / Q000378:metabolism
chemical_list: D017931:DNA Primers; D014409:Tumor Necrosis Factor-alpha; D005437:Flucytosine; D043525:Cytosine Deaminase