Immunocytochemical localization of NOVH protein and ultrastructural characteristics of NCI-H295R cells

J Submicrosc Cytol Pathol. 2001 Jul;33(3):251-60.

Abstract

In this study we present biochemical and immunocytochemical results on NOVH protein secretion and localization in NCI-H295R cells, as well as results on the ultrastructural characteristics of NCI-H295R cells. NCI-H295R cells were characterized by small quantities of rough and smooth endoplasmic reticulum, many free ribosomes, large nuclei with prominent nucleoli, numerous elongated mitochondria, a few Golgi complexes, and a small number of lipid droplets. Large numbers of coated pits and coated vesicles were present, but no secretory granules or exocytotic profiles were seen. Best ultrastructural preservation of NCI-H295R cells was achieved when fixation was done directly on the culture dishes and the cells were detached by scraping. Our biochemical results showed that NCI-H295R cells secreted large amounts of NOVH protein. The immunocytochemical localization of NOVH protein showed that the protein was localized in the cytoplasm, the plasma membrane and the nuclear envelope. This localization pattern, along with the ultrastructural and biochemical findings raise interesting questions on the function(s) and the mode of secretion of NOVH protein.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adrenal Cortex Neoplasms / chemistry
  • Adrenal Cortex Neoplasms / metabolism*
  • Adrenal Cortex Neoplasms / ultrastructure
  • Carcinoma / chemistry
  • Carcinoma / metabolism*
  • Carcinoma / ultrastructure
  • Cell Nucleus / ultrastructure
  • Connective Tissue Growth Factor
  • Fluorescent Antibody Technique, Indirect
  • Growth Substances / analysis
  • Growth Substances / metabolism*
  • Humans
  • Immediate-Early Proteins / analysis
  • Immediate-Early Proteins / metabolism*
  • Immunohistochemistry
  • Intercellular Signaling Peptides and Proteins*
  • Mitogens / analysis
  • Mitogens / metabolism*
  • Nephroblastoma Overexpressed Protein
  • Organelles / ultrastructure
  • Tumor Cells, Cultured

Substances

  • CCN2 protein, human
  • CCN3 protein, human
  • Growth Substances
  • Immediate-Early Proteins
  • Intercellular Signaling Peptides and Proteins
  • Mitogens
  • Nephroblastoma Overexpressed Protein
  • Connective Tissue Growth Factor