Laminin promotes differentiation of NB4 promyelocytic leukemia cells with all-trans retinoic acid

Blood. 1996 Jul 1;88(1):261-7.

Abstract

The promyelocytic leukemia cell line, NB4, carries the t(15; 17) translocation and undergoes limited maturation in response to differentiation agents. Growth on laminin enhanced the ability of all-trans retinoic acid (ATRA) to promote morphologic maturation of these cells. Although exposure to ATRA in suspension yielded minimal maturation beyond the myelocyte stage, after 72 hours of exposure to ATRA on laminin the cells acquired the histologic appearance of metamyelocytes, band forms, and segmented neutrophils. After 96 hours, some cells acquired a spindle shape and became tightly adherent. Growth on collagen types I, III, IV, or fibronectin did not have this effect, although some cells did adhere to fibronectin. NB4 cells treated with ATRA in suspension or on laminin acquired the equivalent ability to reduce nitroblue tetrazolium or cytochrome C. Despite the improved morphologic maturation on laminin, the cells did not express secondary granule proteins such as lactoferrin or neutrophil collagenase. In addition, growth on laminin abolished cell proliferation in the presence of ionomycin. Growth on laminin and/or with ATRA induced new expression of alpha 6 integrin, a laminin receptor, as assessed by reverse transcription-polymerase chain reaction. Different conditions of growth (laminin or differentiation agent) resulted in specific patterns of expression of the alpha 6A and alpha 6B isoforms. Treatment with ATRA also resulted in the acquisition of high-level surface expression of alpha 6 integrin, as assessed by flow cytometry. Thus, treatment of NB4 promyelocytic leukemia cells with ATRA induced expression of alpha 6 integrin (a laminin receptor alpha-chain) and enabled more advanced maturation when the cells were grown on the extracellular matrix component, laminin, compared with tissue culture plastic.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Antigens, CD / biosynthesis
  • Antigens, CD / genetics
  • Base Sequence
  • Calcium / metabolism
  • Cell Culture Techniques / methods
  • Cell Differentiation / drug effects
  • Cell Division / drug effects
  • Collagen / pharmacology
  • Drug Synergism
  • Fibronectins / pharmacology
  • Gene Expression Regulation, Leukemic / drug effects*
  • Humans
  • Integrin alpha6
  • Integrin beta1 / biosynthesis
  • Integrin beta1 / genetics
  • Ionomycin / pharmacology
  • Ionophores / pharmacology
  • Laminin / pharmacology*
  • Leukemia, Promyelocytic, Acute / pathology*
  • Molecular Sequence Data
  • Neoplasm Proteins / biosynthesis
  • Neoplasm Proteins / genetics
  • Neoplastic Stem Cells / drug effects*
  • Neoplastic Stem Cells / pathology
  • Neoplastic Stem Cells / ultrastructure
  • Oxidation-Reduction
  • Polymerase Chain Reaction
  • Receptors, Laminin / biosynthesis
  • Receptors, Laminin / genetics
  • Tretinoin / pharmacology*
  • Tumor Cells, Cultured / drug effects

Substances

  • Antigens, CD
  • Fibronectins
  • Integrin alpha6
  • Integrin beta1
  • Ionophores
  • Laminin
  • Neoplasm Proteins
  • Receptors, Laminin
  • Ionomycin
  • Tretinoin
  • Collagen
  • Calcium