title: [Construction of Lentiviral Vector Over-Expressing MEG3 and Its Effect on XG-7 Cell Apoptosis].
abstract: OBJECTIVE
  To construct a recombinant lentiviral expression vectors carrying MEG3 and to evaluate its effects on XG-7 cell apoptosis.
  
  
  METHODS
  A full-length genomic fragment of human MEG3 was cloned from the pcDNA3.0-MEG3 packaging plasmid and was amplified by PCR. New restriction sites were introduced to be blunted with T4 DNA Ligase. The sequence of the amplified segments was sub-cloned into lentivirus expression vector pCDH-EF1-MCS-T2A-copGFP.The recombined lentiviral expression vector was transfected into 293T cells. FACS was used to detect the effect of MEG3 on XG-7 cell apoptosis after being infected by optimized MOI.
  
  
  RESULTS
  The recombined lentiviral expression vector pCDH-EF1-MEG3-copGFP was constructed successfully. The results showed that pCDH-EF1-MEG3-copGFP could increase the mRNA expression of MEG3 dramatically, its transfection efficiency was more than 90%. The apoptosis rate in XG-7 cells (26.8±2.8%) was very significantly higher than that of the control group (P<0.01).
  
  
  CONCLUSIONS
  The recombined lentiviral LncRNA expression vector targeting MEG3, pCDH-EF1-MEG3-copGFP, has been successfully constructed, the pCDH-EF1-MEG3-copGFP can induce the cell apoptosis in human myeloma cell lines. This study set up a basis to further explore the relationship between human myeloma cells and LncRNA-MEG3 gene.
authors.affiliation: Department of Hematology, The 306 Hospital, The Chinese People's Liberation Army, Beijing 100101,China. E-mail: yikunzhang1307@163.com.
authors.forename: Yi-Kun
authors.initials: YK
authors.lastname: Zhang
authors.affiliation: Institute of Radiation and Irradiation Medicine,Academy of Military Medical Sciences,Beijing 100850,China.
authors.forename: Hua
authors.initials: H
authors.lastname: Wang
authors.affiliation: Institute of Radiation and Irradiation Medicine,Academy of Military Medical Sciences,Beijing 100850,China.
authors.forename: Feng-Jun
authors.initials: FJ
authors.lastname: Xiao
authors.affiliation: Institute of Radiation and Irradiation Medicine,Academy of Military Medical Sciences,Beijing 100850,China.
authors.forename: Xiao-Yan
authors.initials: XY
authors.lastname: Zhang
authors.affiliation: Department of Hematology, The 306 Hospital, The Chinese People's Liberation Army, Beijing 100101,China.
authors.forename: Pei-Lin
authors.initials: PL
authors.lastname: Liu
authors.affiliation: Department of Hematology, The 306 Hospital, The Chinese People's Liberation Army, Beijing 100101,China.
authors.forename: Quan-Xing
authors.initials: QX
authors.lastname: Shi
authors.affiliation: Department of Hematology, The 306 Hospital, The Chinese People's Liberation Army, Beijing 100101,China.
authors.forename: Zhao
authors.initials: Z
authors.lastname: Yin
authors.affiliation: Department of Hematology, The 306 Hospital, The Chinese People's Liberation Army, Beijing 100101,China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Lei
authors.affiliation: Institute of Radiation and Irradiation Medicine,Academy of Military Medical Sciences,Beijing 100850,China. E-mail: wangls@bmi.ac.cn.
authors.forename: Li-Sheng
authors.initials: LS
authors.lastname: Wang
mesh_terms: D017209:Apoptosis; D001483:Base Sequence; D002460:Cell Line; D005822:Genetic Vectors; D006801:Humans; D016086:Lentivirus; D010957:Plasmids; D014162:Transfection