title: ARE/SUZ12 dual specifically-regulated adenoviral TK/GCV system for CML blast crisis cells.
abstract: BACKGROUND
  Treatment of blast phase chronic myeloid leukemia (BP-CML) remains a challenge, and the median survival is less than 6 months. Because effective treatments are lacking, we studied tight targeting of blast crisis CML cells using adenoviral (Ad) vectors expressing a HSV-TK system under dual control of a specific SUZ12 promoter and an antioxidant response element (ARE).
  
  
  METHODS
  A potential SUZ12 promoter fragment was designed with bioinformatics databases and identified with a luciferase assay. Next, we cloned the ARE element of the NQO1 gene and developed Ad vectors expressing TK kinase or luciferase under the dual control of a specific SUZ12 promoter and an ARE element. An in vitro transfection assay with Ad-ARE/SUZ12-Luc was used to determine promoter activity of ARE/SUZ12 regulatory element in blast crisis CML cells. After incubating human BP-CML-derived cells with Ad-ARE/SUZ12-TK and ganciclovir, Western blot, CCK8, Immunofluorescent assays and Annexin V assays were conducted to assess the efficacy of an ARE/SUZ12 dual-specific TK/GCV system for BP-CML cell lines.
  
  
  RESULTS
  Here, luciferase data confirmed significantly higher and specificer promoter activity of the ARE/SUZ12 composite component in CML blast crisis-derived cell lines (K562, KCL22, and K562/G01) compared to HepG2 cells, and Ad-AS-TK/GCV system could exhibit enhanced apoptotic effects and decreased cell viability for BP-CML cell lines. Additionally, Ad-AS-TK/GCV system altered expression of cycle-related and apoptosis-related proteins in BP-CML cell lines.
  
  
  CONCLUSIONS
  Thus, ARE/SUZ12 dual targeting TK/GCV system was effective in killing BP-CML cells. Moreover, efficacy and specificity of CML cell eradication were enhanced by synergistic effects of ARE/SUZ12 dual-specific regulation. We conclude that suicide gene-targeted therapy might hold promise for BP-CML treatment.
authors.affiliation: Department of Clinical Hematology, Key Laboratory of Laboratory Medical Diagnostics of Ministry of Education, Chongqing Medical University, No.1, Yixueyuan Road, Chongqing, 400016, People's Republic of China. zubailing@sina.com.
authors.forename: Bailing
authors.initials: B
authors.lastname: Zu
authors.affiliation: Department of Clinical Laboratory, Shanghai Pudong Hospital, Fudan University Pudong Medical Center, Shanghai, China. shiyi19755@126.com.
authors.forename: Yi
authors.initials: Y
authors.lastname: Shi
authors.affiliation: Department of Clinical Hematology, Key Laboratory of Laboratory Medical Diagnostics of Ministry of Education, Chongqing Medical University, No.1, Yixueyuan Road, Chongqing, 400016, People's Republic of China. 740938405@qq.com.
authors.forename: Min
authors.initials: M
authors.lastname: Xu
authors.affiliation: Department of Clinical Laboratory, Shanghai Children's Medical Center, Shanghai Jiaotong University School of Medicine, Shanghai, China. yuanlaoyoucai@126.com.
authors.forename: Guoling
authors.initials: G
authors.lastname: You
authors.affiliation: Department of Clinical Hematology, Key Laboratory of Laboratory Medical Diagnostics of Ministry of Education, Chongqing Medical University, No.1, Yixueyuan Road, Chongqing, 400016, People's Republic of China. huangzhenlan123@126.com.
authors.forename: Zhenglan
authors.initials: Z
authors.lastname: Huang
authors.affiliation: Department of Clinical Hematology, Key Laboratory of Laboratory Medical Diagnostics of Ministry of Education, Chongqing Medical University, No.1, Yixueyuan Road, Chongqing, 400016, People's Republic of China. 565715711@qq.com.
authors.forename: Miao
authors.initials: M
authors.lastname: Gao
authors.affiliation: Department of Clinical Hematology, Key Laboratory of Laboratory Medical Diagnostics of Ministry of Education, Chongqing Medical University, No.1, Yixueyuan Road, Chongqing, 400016, People's Republic of China. fengwlcqmu@sina.com.
authors.forename: Wenli
authors.initials: W
authors.lastname: Feng
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D001752:Blast Crisis / Q000188:drug therapy / Q000235:genetics / Q000473:pathology; D002453:Cell Cycle / Q000187:drug effects / Q000235:genetics; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D015774:Ganciclovir / Q000494:pharmacology*; D005786:Gene Expression Regulation; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D051267:NF-E2-Related Factor 2 / Q000378:metabolism; D009363:Neoplasm Proteins; D063151:Polycomb Repressive Complex 2 / Q000235:genetics*; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics; D020218:Response Elements / Q000235:genetics*; D013937:Thymidine Kinase / Q000235:genetics*; D014157:Transcription Factors; D015533:Transcriptional Activation; D014161:Transduction, Genetic
chemical_list: D051267:NF-E2-Related Factor 2; D009363:Neoplasm Proteins; D012333:RNA, Messenger; C430461:SUZ12 protein, human; D014157:Transcription Factors; D063151:Polycomb Repressive Complex 2; D013937:Thymidine Kinase; D015774:Ganciclovir