title: Huachansu suppresses human bladder cancer cell growth through the Fas/Fasl and TNF- alpha/TNFR1 pathway in vitro and in vivo.
abstract: BACKGROUND
  Huachansu (HCS), a class of toxic steroids extracted from toad venom, which has been shown to be a valuable anticancer drug in many kinds of cancers. However, the effect of HCS on bladder cancer has not been elucidated. In this study, we focused on the antitumor activities and related mechanisms of HCS on bladder cancer in vitro and in vivo.
  
  
  METHODS
  Cell viability of T24, EJ, RT-4, SV-HUC-1 cells after HCS treatment was measured by MTS, whereas the changes of cell morphology were observed by transmission electron microscopy. The early apoptosis induced by HCS was evaluated by flow cytometry, and the expression level of apoptosis-related molecules (Bax, Bcl-2, XIAP, PARP, cleaved-Caspases 3, 8, 9) was detected using Western blot. We then evaluated the impact of HCS on the expression of Fas/Fasl, TNF- alpha/TNFR1, and the activation of NF-Kappa B pathway, and furthermore the effect of these pathways in HCS induced-apoptosis were also detected. At last, xenograft tumor in nude mice was used to further investigate the antitumor effect of HCS in vivo.
  
  
  RESULTS
  Our results showed that HCS could efficiently inhibit proliferation and induce apoptosis in human bladder cancer cell lines. The expression of Fas, Fasl, TNF- alpha were all elevated at both mRNA and protein level after HCS treatment. Furthermore, down regulation of TNF- alpha, TNFR1, Fas or inhibition of Fas/Fasl interaction decreased the relative number of death cells induced by HCS. In vivo, HCS treatment significantly suppressed tumor growth and induced apoptosis in xenografts tumor in nude mice.
  
  
  CONCLUSIONS
  HCS could efficiently inhibit proliferation and induce apoptosis in human bladder cancer cells in vitro and in vivo, which is largely mediated by Fas/Fasl and TNF- alpha/TNFR1 pathway.
authors.affiliation: Department and Institute of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China. yangtaotao123321@163.com.
authors.forename: Tao
authors.initials: T
authors.lastname: Yang
authors.affiliation: Department and Institute of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China. shanshupai@163.com.
authors.forename: Runlin
authors.initials: R
authors.lastname: Shi
authors.affiliation: Department of Urology, Central Hospital of Wuhan, Wuhan, 430014, China. changlei1025@163.com.
authors.forename: Lei
authors.initials: L
authors.lastname: Chang
authors.affiliation: Department and Institute of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China. 15172407669@163.com.
authors.forename: Kun
authors.initials: K
authors.lastname: Tang
authors.affiliation: Department and Institute of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China. chenke@whu.edu.cn.
authors.forename: Ke
authors.initials: K
authors.lastname: Chen
authors.affiliation: Department and Institute of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China. cdyi1987@126.com.
authors.forename: Gan
authors.initials: G
authors.lastname: Yu
authors.affiliation: Department and Institute of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China. tyf113@gmail.com.
authors.forename: Yuanfeng
authors.initials: Y
authors.lastname: Tian
authors.affiliation: Department of Urology, Central Hospital of Wuhan, Wuhan, 430014, China. guoYL111@aliyun.com.
authors.forename: Yonglian
authors.initials: Y
authors.lastname: Guo
authors.affiliation: Department and Institute of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China. hewei_6688@163.com.
authors.forename: Wei
authors.initials: W
authors.lastname: He
authors.affiliation: Department and Institute of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China. songxdd@126.com.
authors.forename: Xiaodong
authors.initials: X
authors.lastname: Song
authors.affiliation: Department and Institute of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China. xuhuawhu@163.com.
authors.forename: Hua
authors.initials: H
authors.lastname: Xu
authors.affiliation: Department and Institute of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China. zhangqun_ye@163.com.
authors.forename: Zhangqun
authors.initials: Z
authors.lastname: Ye
mesh_terms: D000664:Amphibian Venoms / Q000494:pharmacology*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D004195:Disease Models, Animal; D053222:Fas Ligand Protein / Q000378:metabolism*; D064593:Heterografts; D006801:Humans; D016328:NF-kappa B / Q000378:metabolism; D047888:Receptors, Tumor Necrosis Factor, Type I / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects*; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism*; D001749:Urinary Bladder Neoplasms / Q000378:metabolism* / Q000473:pathology; D019014:fas Receptor / Q000378:metabolism*
chemical_list: D000664:Amphibian Venoms; D053222:Fas Ligand Protein; D016328:NF-kappa B; D047888:Receptors, Tumor Necrosis Factor, Type I; D014409:Tumor Necrosis Factor-alpha; D019014:fas Receptor; C545813:huachansu