title: [Construction of a GFP/Puro double-labeled lentiviral vector containing CK8 interfering RNA and its effect on cell apoptosis in vitro].
abstract: OBJECTIVE
  To construct a GFP/Puro double-labeled lentiviral expression vector for CK8 silencing and assess the effects of CK8 silencing on cell apoptosis.
  
  
  METHODS
  The siRNA sequences of CK8 were inserted into the lentiviral expression vector GV248 and transfected into 293T cells with the packaging plasmids PMD and SPA. The lentivirus was collected at 24 and 36 h post-transfection. Flow cytometry was used to detect the virus titer and the positive cells were selected with puromycin. The knockdown of CK8 was examined by Western blotting. The effect of CK8 down-regulation on cell apoptosis induced by cisplatin was detected with Annexin V/PI staining.
  
  
  CONCLUSIONS
  We successfully constructed CK8 interference lentiviral vector and obtained a stable cell line with CK8 knock-down that was sensitive to cisplatin-induced apoptosis.
authors.affiliation: Department of Pharmaceutical Engineering, Tianjin University, Tianjin 300072, China. E-mail: 369king@163.com.
authors.forename: Yanchao
authors.initials: Y
authors.lastname: Jin
authors.forename: Ronghua
authors.initials: R
authors.lastname: Yin
authors.forename: Weiwei
authors.initials: W
authors.lastname: Zheng
authors.forename: Xiangzhen
authors.initials: X
authors.lastname: Kong
authors.forename: Yiqun
authors.initials: Y
authors.lastname: Zhan
authors.forename: Xiaoming
authors.initials: X
authors.lastname: Yang
authors.forename: Changyan
authors.initials: C
authors.lastname: Li
mesh_terms: D017209:Apoptosis; D002460:Cell Line; D015536:Down-Regulation; D005822:Genetic Vectors; D006801:Humans; D053533:Keratin-8 / Q000235:genetics*; D016086:Lentivirus; D010957:Plasmids; D034622:RNA Interference; D034741:RNA, Small Interfering; D014162:Transfection
chemical_list: D053533:Keratin-8; D034741:RNA, Small Interfering