title: [Construction of a CD147 lentiviral expression vector and establishment of its stably transfected A549 cell line].
abstract: OBJECTIVE
  CD147, a type of transmembrane glycoprotein embedded on the surface of tumor cells, can promote tumor invasion and metastasis. This aim of this study is to construct a CD147 lentiviral expression vector, establish its stably transfected A549 cell line, and observe the effect of CD147 on MMP-9 proliferation as well as on the invasive ability of human lung adenocarcinoma cells.
  
  
  METHODS
  Full-length CD147 gene was amplified by real-time polymerase chain reaction (RT-PCR), inserted into a pEGFP vector to construct pEGFP-CD147 and pEGFP vectors, and then transfected into 293FT cells to precede the lentivirus equipment package. Subsequently, we collected the lentivirus venom to infect the A549 cells and establish a stable, overexpressed cell line named A549-CD147. The mRNA expression of MMP-9 was examined by RT-PCR. The proliferation and invasive ability of the human lung cancer cells before and after transfection were examined by the CCK-8 and Transwell methods.
  
  
  RESULTS
  A CD147 lentiviral expression vector (pEGFP-CD147) was successfully constructed by restrictive enzyme digestion and plasmid sequencing. RT-PCR and Western blot analyses revealed increased mRNA and protein expression of CD147 gene in cells transfected with pEGFP-CD147 compared with the control groups. Therefore, the A549-CD147 cell line was successfully established through the experiment. The mRNA expression of MMP-9 also significantly increased after the upregulation of CD147 expression. Meanwhile, CCK-8 and Transwell assays indicated that the proliferation and invasive ability significantly increased in the A549-CD147 cells.
  
  
  CONCLUSIONS
  A lentiviral CD147 expression vector and its A549 cell line (A549-CD14) were successfully constructed. CD147 overexpression upregulated the protein expression of MMP-9, and strengthened the proliferation and invasive ability of human lung adenocarcinoma cells.
authors.affiliation: Department of Pulmonary Oncology, Affiliated Hospital of Academy of Military Medical Sciences, Beijing 100071, China.
authors.forename: Shaoxing
authors.initials: S
authors.lastname: Yang
authors.forename: Chuanhao
authors.initials: C
authors.lastname: Tang
authors.forename: Sihan
authors.initials: S
authors.lastname: Wang
authors.forename: Santai
authors.initials: S
authors.lastname: Song
authors.forename: Xiaoqing
authors.initials: X
authors.lastname: Liu
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D051926:Basigin / Q000235:genetics* / Q000378:metabolism; D015153:Blotting, Western; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000235:genetics; D049109:Cell Proliferation; D015972:Gene Expression Regulation, Neoplastic; D005822:Genetic Vectors / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D008175:Lung Neoplasms / Q000235:genetics / Q000473:pathology; D020780:Matrix Metalloproteinase 9 / Q000235:genetics / Q000378:metabolism; D008856:Microscopy, Fluorescence; D009361:Neoplasm Invasiveness; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection / Q000379:methods*; D015854:Up-Regulation
chemical_list: C497886:BSG protein, human; D051926:Basigin; D049452:Green Fluorescent Proteins; D020780:Matrix Metalloproteinase 9