title: [Establishment of a human bladder cancer cell line stably co-expressing hSPRY2 and luciferase genes and its subcutaneous tumor xenograft model in nude mice].
abstract: Objective To establish a human bladder cancer cell line stably co-expressing human sprouty2 (hSPRY2) and luciferase (Luc) genes simultaneously, and develop its subcutaneous tumor xenograft model in nude mice. Methods The hSPRY2 and Luc gene segments were amplified by PCR, and were cloned into lentiviral vector pCDH and pLVX respectively to produce corresponding lentivirus particles. The J82 human bladder cancer cells were infected with these two kinds of lentivirus particles, and then further screened by puromycin and G418. The expressions of hSPRY2 and Luc genes were detected by bioluminescence, immunofluorescence and Western blot analysis. The screened J82-hSPRY2/Luc cells were injected subcutaneously into BALB/c nude mice, and the growth of tumor was monitored dynamically using in vivo fluorescence imaging system. Results J82-hSPRY2/Luc cell line stably expressing hSPRY2 and Luc genes was established successfully. Bioluminescence, immunofluorescence and Western blot analysis validated the expressions of hSPRY2 and Luc genes. The in vivo fluorescence imaging system showed obvious fluorescence in subcutaneous tumor xenograft in nude mice. Conclusion The J82-hSPRY2/Luc bladder cancer cell line and its subcutaneous tumor xenograft model in nude mice have been established successfully.
authors.affiliation: Department of Urology, General Hospital of PLA, Beijing 100853, China.
authors.forename: Xiaotao
authors.initials: X
authors.lastname: Yin
authors.affiliation: Department of Urology, Bethune International Peace Hospital of PLA, Shijiazhuang 050081, China.
authors.forename: Fanglong
authors.initials: F
authors.lastname: Li
authors.affiliation: Department of Urology, General Hospital of PLA, Beijing 100853, China.
authors.forename: Yipeng
authors.initials: Y
authors.lastname: Jin
authors.affiliation: Department of Urology, General Hospital of PLA, Beijing 100853, China.
authors.forename: Zhaoyang
authors.initials: Z
authors.lastname: Yin
authors.affiliation: Department of Urology, General Hospital of PLA, Beijing 100853, China.
authors.forename: Siyong
authors.initials: S
authors.lastname: Qi
authors.affiliation: Institute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100850, China.
authors.forename: Shuai
authors.initials: S
authors.lastname: Wu
authors.affiliation: Institute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100850, China.
authors.forename: Zicheng
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Department of Internal Medicine, PLA 316 Hospital, Beijing 100093, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Wang
authors.affiliation: Institute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100850, China. *Corresponding authors, E-mail: yujyun@126.com.
authors.forename: Jiyun
authors.initials: J
authors.lastname: Yu
authors.affiliation: Department of Urology, General Hospital of PLA, Beijing 100853, China. *Corresponding authors, E-mail: jpgao@163.com.
authors.forename: Jiangping
authors.initials: J
authors.lastname: Gao
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D017930:Genes, Reporter; D064593:Heterografts; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics* / Q000378:metabolism; D008156:Luciferases / Q000235:genetics* / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D001749:Urinary Bladder Neoplasms / Q000235:genetics* / Q000378:metabolism
chemical_list: D047908:Intracellular Signaling Peptides and Proteins; D008565:Membrane Proteins; C485561:SPRY2 protein, human; D008156:Luciferases