title: Anti-tumor function of double-promoter regulated adenovirus carrying SEA gene, in the treatment of bladder cancer.
abstract: To construct an adenovirus carrying SEA gene and regulated by telomerase reverse transcriptase (hTERT) and hypoxia-inducible factor (HIF) promoters and investigate its anti-tumor function in vitro, as well as its role in lymphocyte production. hTERT and HIF genes were cloned into adenovirus E1A and E1B shuttle plasmids. The control vector for SEA gene expression is under the regulation of CMV and SV40 promoters. Double regulation was obtained through homologous recombination. The positive clones of replicable adenovirus H2-SEA-Ad were selected by plaque assay. The adenovirus was purified, titrated, and DNA was verified by PCR. The obtained virus was used to infect EJ bladder tumor cells and the SEA Mrna, and protein expression was measured by RT-PCR, western blot, and immunofluorescence microscopy, respectively. Co-culture of lymphocytes and tumor cells was observed dynamically under microscope. The construction of shuttle plasmid p315-CSS-SEA was confirmed by PCR and DNA sequencing. Insertion of superantigen SEA gene in adenovirus (H2-SEA-Ad.SEA) was obtained by homologous recombination. In lymphocytes and tumor cell co-culture, the number of viable tumor cells in test groups was significantly lower than that in control group after 12, 24, and 48 h of treatment. Production of interleukin-2, interleukin-4, and tumor necrosis factor were higher in test groups than in control group. Expression of SEA gene in bladder tumor cells by adenoviral vector caused reduced tumor cell proliferation, as well as stimulation of inflammatory cytokine productions in co-cultures with lymphocytes.
authors: Department of Urology, People's Hospital Affiliated to Jiangsu University, Zhenjiang 212002, China. hujianpeng9909@yahoo.cn
authors: Jianpeng
authors: J
authors: Hu
authors: Xujun
authors: X
authors: Xuan
authors: Conghui
authors: C
authors: Han
authors: Lin
authors: L
authors: Hao
authors: Peiying
authors: P
authors: Zhang
authors: Meng
authors: M
authors: Chen
authors: Houguang
authors: H
authors: He
authors: Tao
authors: T
authors: Fan
authors: Binzheng
authors: B
authors: Dong
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D018920:Coculture Techniques; D003587:Cytomegalovirus / Q000235:genetics; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D051793:Hypoxia-Inducible Factor 1 / Q000235:genetics / Q000378:metabolism; D007376:Interleukin-2 / Q000378:metabolism; D015847:Interleukin-4 / Q000378:metabolism; D008214:Lymphocytes / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D011401:Promoter Regions, Genetic; D013539:Simian virus 40 / Q000235:genetics; D018089:Superantigens / Q000235:genetics* / Q000378:metabolism; D019098:Telomerase / Q000235:genetics / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D001749:Urinary Bladder Neoplasms / Q000473:pathology
chemical_list: D051793:Hypoxia-Inducible Factor 1; D007376:Interleukin-2; D018089:Superantigens; D014409:Tumor Necrosis Factor-alpha; D015847:Interleukin-4; C509186:TERT protein, human; D019098:Telomerase