title: Synthetic Bax-Anti Bcl2 combination module actuated by super artificial hTERT promoter selectively inhibits malignant phenotypes of bladder cancer.
abstract: BACKGROUND
  The synthetic biology technology which enhances the specificity and efficacy of treatment is a novel try in biomedical therapy during recent years. A high frequency of somatic mutations was shown in the human telomerase reverse transcriptase (hTERT) promoter in bladder cancer, indicating that a mutational hTERT promoter might be a tumor-specific element for bladder cancer therapy. In our study, we aimed to construct a synthetic combination module driven by a super artificial hTERT promoter and to investigate its influence on the malignant phenotypes of bladder cancer.
  
  
  METHODS
  The dual luciferase assay system was used to verify the driven efficiency and tumor-specificity of the artificial hTERT promoter and to confirm the relationship between ETS-1 and the driven efficiency of the artificial hTERT promoter. CCK-8 assay and MTT assay were used to test the effects of the Bax-Anti Bcl2 combination module driven by the artificial hTERT promoter on cell proliferation. Simultaneously, the cell apoptosis was detected by the caspase 3ELISA assay and the flow cytometry analysis after transfection. The results of CCK-8 assay and MTT assay were analyzed by ANOVA. The independent samples t-test was used to analyze other data.
  
  
  RESULTS
  We demonstrated that the artificial hTERT promoter had a higher driven efficiency which might be regulated by transcription factor ETS-1 in bladder cancer cells, compared with wild-type hTERT promoter. Meanwhile, the artificial hTERT promoter showed a strong tumor-specific effect. The cell proliferation inhibition and apoptosis induction were observed in artificial hTERT promoter- Bax-Anti Bcl2 combination module -transfected bladder cancer 5637 and T24 cells, but not in the module -transfected normal human fibroblasts.
  
  
  CONCLUSIONS
  This module offers us a useful synthetic biology platform to inhibit the malignant phenotypes of bladder cancer in a more specific and effective way.
authors.affiliation: Key Laboratory of Medical Reprogramming Technology, Shenzhen Second People's Hospital, The First Affiliated Hospital of Shenzhen University, Shenzhen, China. sumc_liuli2013@163.com.|Shantou University Medical College, Shantou, China. sumc_liuli2013@163.com.
authors.forename: Li
authors.initials: L
authors.lastname: Liu
authors.affiliation: Key Laboratory of Medical Reprogramming Technology, Shenzhen Second People's Hospital, The First Affiliated Hospital of Shenzhen University, Shenzhen, China. liuyuchenmdcg@163.com.
authors.forename: Yuchen
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Urology Department, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China. zhangtianbiao215@126.com.
authors.forename: Tianbiao
authors.initials: T
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Medical Reprogramming Technology, Shenzhen Second People's Hospital, The First Affiliated Hospital of Shenzhen University, Shenzhen, China. rayman_wu@126.com.
authors.forename: Hanwei
authors.initials: H
authors.lastname: Wu
authors.affiliation: Key Laboratory of Medical Reprogramming Technology, Shenzhen Second People's Hospital, The First Affiliated Hospital of Shenzhen University, Shenzhen, China. muk_lin@163.com.
authors.forename: Muqi
authors.initials: M
authors.lastname: Lin
authors.affiliation: Urology Department, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China. wangchaoliang2008@yahoo.com.
authors.forename: Chaoliang
authors.initials: C
authors.lastname: Wang
authors.affiliation: Key Laboratory of Medical Reprogramming Technology, Shenzhen Second People's Hospital, The First Affiliated Hospital of Shenzhen University, Shenzhen, China. yonghao_zhan@163.com.|Shantou University Medical College, Shantou, China. yonghao_zhan@163.com.
authors.forename: Yonghao
authors.initials: Y
authors.lastname: Zhan
authors.affiliation: Key Laboratory of Medical Reprogramming Technology, Shenzhen Second People's Hospital, The First Affiliated Hospital of Shenzhen University, Shenzhen, China. zqboy008@163.com.
authors.forename: Qing
authors.initials: Q
authors.lastname: Zhou
authors.affiliation: Urology Department, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China. zdyfynksl@126.com.
authors.forename: Baoping
authors.initials: B
authors.lastname: Qiao
authors.affiliation: Key Laboratory of Medical Reprogramming Technology, Shenzhen Second People's Hospital, The First Affiliated Hospital of Shenzhen University, Shenzhen, China. xiaojuan26@gmail.com.
authors.forename: Xiaojuan
authors.initials: X
authors.lastname: Sun
authors.affiliation: Key Laboratory of Medical Reprogramming Technology, Shenzhen Second People's Hospital, The First Affiliated Hospital of Shenzhen University, Shenzhen, China. qiaoxiazhang@163.com.
authors.forename: Qiaoxia
authors.initials: Q
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Medical Reprogramming Technology, Shenzhen Second People's Hospital, The First Affiliated Hospital of Shenzhen University, Shenzhen, China. xiaoqiangguo123@163.com.
authors.forename: Xiaoqiang
authors.initials: X
authors.lastname: Guo
authors.affiliation: Key Laboratory of Medical Reprogramming Technology, Shenzhen Second People's Hospital, The First Affiliated Hospital of Shenzhen University, Shenzhen, China.|Shanghai-MOST Key Laboratory of Health and Disease Genomics, Chinese National Human Genome Center at Shanghai, Shanghai, China.
authors.forename: Guoping
authors.initials: G
authors.lastname: Zhao
authors.affiliation: Urology Department, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China. zhangweixing1957@yahoo.com.
authors.forename: Weixing
authors.initials: W
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Medical Reprogramming Technology, Shenzhen Second People's Hospital, The First Affiliated Hospital of Shenzhen University, Shenzhen, China. pony8980@163.com.|Department of Urology, Peking University First Hospital, Institute of Urology, Peking University, National Urological Cancer Centre, Beijing, 100034, China. pony8980@163.com.
authors.forename: Weiren
authors.initials: W
authors.lastname: Huang
mesh_terms: D045744:Cell Line, Tumor; D049109:Cell Proliferation; D015972:Gene Expression Regulation, Neoplastic; D015316:Genetic Therapy; D006801:Humans; D010641:Phenotype; D011401:Promoter Regions, Genetic; D050784:Proto-Oncogene Protein c-ets-1; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000037:antagonists & inhibitors* / Q000235:genetics; D034741:RNA, Small Interfering / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000494:pharmacology; D058615:Synthetic Biology; D019098:Telomerase / Q000235:genetics*; D001749:Urinary Bladder Neoplasms / Q000235:genetics* / Q000628:therapy; D051028:bcl-2-Associated X Protein / Q000235:genetics*
chemical_list: C494826:BAX protein, human; C000595707:BCL2 protein, human; C494041:ETS1 protein, human; D050784:Proto-Oncogene Protein c-ets-1; D019253:Proto-Oncogene Proteins c-bcl-2; D034741:RNA, Small Interfering; D011994:Recombinant Proteins; D051028:bcl-2-Associated X Protein; C509186:TERT protein, human; D019098:Telomerase