Establishment and characterization of a human adrenocortical carcinoma cell line that expresses multiple pathways of steroid biosynthesis

Adi F. Gazdar, Herbert K. Oie, Cedric H. Shackleton, T. R. Chen, Timothy J. Triche, Charles E. Myers, George P. Chrousos, Murray F. Brennan, C. A. Stein, Renato V. La Rocca

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Abstract

We established a continuous cell line, NCI-H295, from an invasive primary adrenocortical carcinoma. The cell line was established in a fully defined medium (HITES) and later could be adapted for growth in a simple medium supplemented only with selenium, insulin, and transferrin and devoid of serum, steroids, fibroblast growth factor, and a source of exogenous cholesterol. NCI-H295 cells had a relatively long population doubling time and were tumorigenic when inoculated s.c. into athymic nude mice. The cultured cells had ultrastructural features of steroidsecreting cells and contained complex cytogenetic abnormalities including the presence of multiple marker chromosomes. Steroid analyses (radioimmunoassays and mass spectrometry), performed 7 to 9 years after culture initiation, demonstrated secretion of more than 30 steroids characteristic of adrenocortical cells. Total unconjugated steroid secretion in serum-supplemented medium was 2.83 μg/106 cells/24 h and about 4-fold less in serum-free medium. The major pathway of pregnenolone metabolism in NCI-H295 cells is androgen synthesis, with formation of dehydroepiandrosterone, androstenedione, testotesterone, and at least three sulfated androgens, as well as estrogens. In addition, formation of cortisol, corticosterone, aldosterone, and 11β-hydroxyandrostenidione indicated the presence of 11β-hydroxylase. Thus, multiple pathways of steroidogenesis are expressed by NCI-H295 cells, including formation of corticosteroids, mineralocorticoids, androgens, and estrogens. Our findings indicate the presence in NCI-H295 cells of all of the major adrenocortical enzyme systems, including 11β-hydroxylase, desmolase, 21α-hydroxylase, 17α-hydroxylase, 18-hydroxylase, lyase, sulfokinase, and aromatase. The NCI-H295 cell line should prove of value in studying the regulation, metabolic pathways, and enzymes involved in steroid formation and secretion. In addition, it may provide insights into the biology and treatment of adrenocortical carcinoma.

Original languageEnglish (US)
Pages (from-to)5488-5496
Number of pages9
JournalCancer Research
Volume50
Issue number17
StatePublished - Sep 1 1990

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Adrenocortical Carcinoma
Steroids
Cell Line
Mixed Function Oxygenases
Androgens
Lyases
Nude Mice
Estrogens
Arylsulfotransferase
Steroid 21-Hydroxylase
Pregnenolone
Mineralocorticoids
Aromatase
Dehydroepiandrosterone
Fibroblast Growth Factors
Androstenedione
Serum-Free Culture Media
Enzymes
Transferrin
Selenium

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Gazdar, A. F., Oie, H. K., Shackleton, C. H., Chen, T. R., Triche, T. J., Myers, C. E., ... La Rocca, R. V. (1990). Establishment and characterization of a human adrenocortical carcinoma cell line that expresses multiple pathways of steroid biosynthesis. Cancer Research, 50(17), 5488-5496.

Establishment and characterization of a human adrenocortical carcinoma cell line that expresses multiple pathways of steroid biosynthesis. / Gazdar, Adi F.; Oie, Herbert K.; Shackleton, Cedric H.; Chen, T. R.; Triche, Timothy J.; Myers, Charles E.; Chrousos, George P.; Brennan, Murray F.; Stein, C. A.; La Rocca, Renato V.

In: Cancer Research, Vol. 50, No. 17, 01.09.1990, p. 5488-5496.

Research output: Contribution to journalArticle

Gazdar, AF, Oie, HK, Shackleton, CH, Chen, TR, Triche, TJ, Myers, CE, Chrousos, GP, Brennan, MF, Stein, CA & La Rocca, RV 1990, 'Establishment and characterization of a human adrenocortical carcinoma cell line that expresses multiple pathways of steroid biosynthesis', Cancer Research, vol. 50, no. 17, pp. 5488-5496.
Gazdar, Adi F. ; Oie, Herbert K. ; Shackleton, Cedric H. ; Chen, T. R. ; Triche, Timothy J. ; Myers, Charles E. ; Chrousos, George P. ; Brennan, Murray F. ; Stein, C. A. ; La Rocca, Renato V. / Establishment and characterization of a human adrenocortical carcinoma cell line that expresses multiple pathways of steroid biosynthesis. In: Cancer Research. 1990 ; Vol. 50, No. 17. pp. 5488-5496.
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abstract = "We established a continuous cell line, NCI-H295, from an invasive primary adrenocortical carcinoma. The cell line was established in a fully defined medium (HITES) and later could be adapted for growth in a simple medium supplemented only with selenium, insulin, and transferrin and devoid of serum, steroids, fibroblast growth factor, and a source of exogenous cholesterol. NCI-H295 cells had a relatively long population doubling time and were tumorigenic when inoculated s.c. into athymic nude mice. The cultured cells had ultrastructural features of steroidsecreting cells and contained complex cytogenetic abnormalities including the presence of multiple marker chromosomes. Steroid analyses (radioimmunoassays and mass spectrometry), performed 7 to 9 years after culture initiation, demonstrated secretion of more than 30 steroids characteristic of adrenocortical cells. Total unconjugated steroid secretion in serum-supplemented medium was 2.83 μg/106 cells/24 h and about 4-fold less in serum-free medium. The major pathway of pregnenolone metabolism in NCI-H295 cells is androgen synthesis, with formation of dehydroepiandrosterone, androstenedione, testotesterone, and at least three sulfated androgens, as well as estrogens. In addition, formation of cortisol, corticosterone, aldosterone, and 11β-hydroxyandrostenidione indicated the presence of 11β-hydroxylase. Thus, multiple pathways of steroidogenesis are expressed by NCI-H295 cells, including formation of corticosteroids, mineralocorticoids, androgens, and estrogens. Our findings indicate the presence in NCI-H295 cells of all of the major adrenocortical enzyme systems, including 11β-hydroxylase, desmolase, 21α-hydroxylase, 17α-hydroxylase, 18-hydroxylase, lyase, sulfokinase, and aromatase. The NCI-H295 cell line should prove of value in studying the regulation, metabolic pathways, and enzymes involved in steroid formation and secretion. In addition, it may provide insights into the biology and treatment of adrenocortical carcinoma.",
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AU - Oie, Herbert K.

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AU - Chen, T. R.

AU - Triche, Timothy J.

AU - Myers, Charles E.

AU - Chrousos, George P.

AU - Brennan, Murray F.

AU - Stein, C. A.

AU - La Rocca, Renato V.

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