Establishment and characterization of a squamous cell carcinoma cell line, designated hZK-1, derived from a metastatic lymph node tumor of the tongue

Hum Cell. 2017 Oct;30(4):319-326. doi: 10.1007/s13577-017-0165-y. Epub 2017 Mar 4.

Abstract

The hZK-1 cell line was successfully established from the metastatic foci of a lymph node of an 82-year-old Japanese woman with squamous cell carcinoma of the tongue. The pathological diagnosis of the tumor was moderately to well-differentiated squamous cell carcinoma. The hZK-1 cells were angular in shape, and had neoplastic and pleomorphic features. Adjacent hZK-1 cells were joined by desmosomes and well-developed microvilli, and many free ribosomes were observed in the cytoplasm. The doubling time of the hZK-1 cells was approximately 36, 33, and 29 h at the 10th, 20th, and 30th passages, respectively. The cell line was shown to be triploid, with a chromosomal distribution of 75-80. Immunocytochemical staining of the hZK-1 cells revealed cytokeratin (CK) 17-, Ki67-, and p53-positive staining, and negative staining for CK13. The hZK-1 cells were negative for human papillomavirus (HPV)-16 or-18 infection. Grafting was not successful when the hZK-1 cells were transplanted into the subcutis of SCID mice. The hZK-1 cells (2 × 106 cells/3 ml of growth medium) secreted vascular endothelial growth factor (VEGF) that reached a concentration of 2.6 ng/ml media after 3 days of culture. Hypoxia enhanced cellular HIF-1α expression and VEGF secretion in hZK-1 cells. The HIF-1α inhibitor YC-1 partially inhibited hypoxia-induced VEGF secretion in ZK-1 cells. The reverse transcription-polymerase chain reaction (RT-PCR) results revealed that the expression of CK17, Ki67, and p53 was elevated in the hZK-1 cells. hZK-1 cells were not sensitive to CDDP, TXT, 5-FU, or a mixture of these three anti-tumor agents.

Keywords: Carcinoma cell; Lymph node; Tumor of the tongue; hZK-1.

MeSH terms

  • Aged, 80 and over
  • Animals
  • Antineoplastic Agents / pharmacology
  • Carcinoma, Squamous Cell / genetics
  • Carcinoma, Squamous Cell / metabolism
  • Carcinoma, Squamous Cell / pathology*
  • Cell Line, Tumor
  • Cell Proliferation
  • Drug Resistance, Neoplasm
  • Female
  • Humans
  • Hypoxia-Inducible Factor 1, alpha Subunit / antagonists & inhibitors
  • Hypoxia-Inducible Factor 1, alpha Subunit / metabolism
  • Indazoles / pharmacology
  • Keratin-13 / metabolism
  • Keratin-17 / metabolism
  • Ki-67 Antigen / metabolism
  • Lymphatic Metastasis
  • Mice, SCID
  • Tongue Neoplasms / genetics
  • Tongue Neoplasms / metabolism
  • Tongue Neoplasms / pathology*
  • Tumor Suppressor Protein p53 / metabolism
  • Vascular Endothelial Growth Factor A / metabolism

Substances

  • Antineoplastic Agents
  • HIF1A protein, human
  • Hypoxia-Inducible Factor 1, alpha Subunit
  • Indazoles
  • Keratin-13
  • Keratin-17
  • Ki-67 Antigen
  • Tumor Suppressor Protein p53
  • Vascular Endothelial Growth Factor A
  • 3-(5'-hydroxymethyl-2'-furyl)-1-benzylindazole