title: [Overexpression of Smac gene enhanced chemotherapeutic sensitivity of esophageal cancer cell line Eca109 to cisplatin].
abstract: OBJECTIVE
  To investigate the effects of Smac gene overexpression on chemotherapeutic sensitivity of esophageal cancer cell line Eca109 to cisplatin.
  
  
  METHODS
  pcDNA3.1-Smac with GFP and pcDNA3.1-Smac with GFP were transfected into esophageal cancer cell line Eca109 by liposome and incubated with G418 for subclone selection.The efficiency of transfection was observed under fluorescence microscope, cellular Smac gene expression were determined  by  Western blot. Cisplatin treated group (1, 5, 10 mg/L) and cisplatin untreated group were selected to treat untransfected and transfected esophageal cancer cell line Eca109.   Apoptosis was determined by Annexin V/PI.
  
  
  RESULTS
  The subclone esophageal cancer cell line Eca109,  stable expressing Smac+GFP and neo+GFP respectively,  were successfully selected,  named as Eca109/Smac,  Eca109/neo. Compared with the Eca109/neo and Eca109,  the Smac expression level of Eca109/Smac was significantly increased(P<0.05). In cisplatin untreated group,  the apoptosis rate was not associated with Smac expression. In cisplatin treated group (1, 5, 10 mg/L),  compared with Eca109/neo and Eca109,  the apoptosis rate of the Eca109/Smac was significantly increased after the treatments of cisplatin,  the difference were significant(P<0.05). In addition,  the apoptosis rate of Eca109/Smac was significantly increased with the concentration of cispaltin increased(P<0.05). The difference between Eca109/neo and Eca109 were not significant.
  
  
  CONCLUSIONS
  Smac gene didn't induce apoptosis in the cisplatin untreated Eca109 cells. Smac gene overexpression could increase chemotherapeutic sensitivity of esophageal cancer cell line Eca109 to cisplatin.
authors.affiliation: Department of Oncosurgery,  First Affiliated Hospital of Medical College of  Xi'an Jiaotong University,  Xi'an 710061,  China.
authors.forename: Ning
authors.initials: N
authors.lastname: Du
authors.forename: Bin
authors.initials: B
authors.lastname: Yang
authors.forename: Li-Juan
authors.initials: LJ
authors.lastname: Hu
authors.forename: Yang
authors.initials: Y
authors.lastname: Zhao
authors.forename: Xin
authors.initials: X
authors.lastname: Sun
authors.forename: Zhi-Wei
authors.initials: ZW
authors.lastname: Guo
authors.forename: Hong
authors.initials: H
authors.lastname: Ren
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D051017:Apoptosis Regulatory Proteins; D015153:Blotting, Western; D045744:Cell Line, Tumor; D002945:Cisplatin / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004938:Esophageal Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D005434:Flow Cytometry; D015972:Gene Expression Regulation, Neoplastic; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D024101:Mitochondrial Proteins / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014162:Transfection
chemical_list: D000970:Antineoplastic Agents; D051017:Apoptosis Regulatory Proteins; C412403:DIABLO protein, human; D047908:Intracellular Signaling Peptides and Proteins; D024101:Mitochondrial Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D002945:Cisplatin