Expression of LAPTM4B in gallbladder carcinoma cells: the role in invasive potential

Hepatogastroenterology. 2010 Mar-Apr;57(98):207-11.

Abstract

Background/aims: It was previously established that LAPTM4B-35 highly expressed in gallbladder carcinoma and being of clinicopathological and prognostic significances. However, expression of LAPTM4B gene in gallbladder carcinoma (GBC-SD), a gallbladder carcinoma cell line, and its role in invasive potential remain unclear.

Methodology: Expression of LAPTM4B in GBC-SD cells was first detected. Plasmids, pcDNA3-AE (containing complete open reading frame of LAPTM4B) and Mock (pcDNA3), were transiently transfected into GBC-SD cells. Invasive phenotypes (migration and invasion) and relative molecules were then shown by transwell assay, crossing river test and Western blot analysis.

Results: Immunocytochemical staining revealed that LAPTM4B-35 positively expressed in cytoplasm of GBC-SD cells. But LAPTM4B-35 expression was obviously weaker in GBC-SD cells than that in BEL-7402 cells (positive control). Besides, cells transfected with pcDNA3-AE presented shorter crossing river time, less migrated and invaded cell numbers, compared with cells transfected with the Mock plasmid and parent cells. Finally, increased expressions of active uPA, MMP-9, pro MMP-2 and active MMP-2 were also observed in cells transfected with pcDNA3-AE.

Conclusions: Our data suggested that LAPTM4B expressed in GBC-SD cells at a relatively low level. Forced overexpression of LAPTM4B increased invasive potential of GBC-SD cells, through modulating molecules associated with degradation of extracellular matrix.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blotting, Western
  • Cell Line, Tumor
  • Cell Movement
  • Dinucleoside Phosphates / metabolism
  • Extracellular Matrix / metabolism
  • Gallbladder Neoplasms / genetics*
  • Gallbladder Neoplasms / metabolism
  • Humans
  • Immunoenzyme Techniques
  • Matrix Metalloproteinase 2 / metabolism
  • Matrix Metalloproteinase 9 / metabolism
  • Membrane Proteins / genetics*
  • Membrane Proteins / metabolism
  • Oncogene Proteins / genetics*
  • Oncogene Proteins / metabolism
  • Open Reading Frames
  • Phenotype
  • Plasmids
  • Statistics, Nonparametric
  • Transfection
  • Tumor Cells, Cultured

Substances

  • Dinucleoside Phosphates
  • LAPTM4B protein, human
  • Membrane Proteins
  • Oncogene Proteins
  • uridylyl-(3'-5')-adenosine
  • MMP2 protein, human
  • Matrix Metalloproteinase 2
  • Matrix Metalloproteinase 9