Establishment and characterization of a human adrenocortical carcinoma cell line that expresses multiple pathways of steroid biosynthesis.
Establishment and characterization of a human adrenocortical carcinoma cell line that expresses multiple pathways of steroid biosynthesis.
复制标题
DOI：
--
复制
发表时间：
1990-09
期刊：
影响因子：
11.2
通讯作者：
A. Gazdar;H. Oie;C. Shackleton;T. Chen;T. Triche;T. Triche;C. Myers;G. Chrousos;G. Chrousos
中科院分区：
文献类型：
--
作者：
A. Gazdar;H. Oie;C. Shackleton;T. Chen;T. Triche;T. Triche;C. Myers;G. Chrousos;G. Chrousos
我们从侵入性的原发性肾上腺皮质癌中建立了连续的细胞系NCI-H295。细胞系在完全定义的培养基中（hITE）建立，后来可以在仅补充硒，胰岛素和转铁蛋白和无血清，类固醇，成纤维细胞生长因子和外源性胆固醇的来源的简单培养基中进行调整，以生长。 。 NCI-H295细胞的种群倍增时间相对较长，当接种S.C时是肿瘤的。进入裸裸鼠。培养的细胞具有类固醇分泌细胞的超微结构特征，并包含复杂的细胞遗传学异常，包括存在多个标记物染色体。培养开始后7至9年进行了类固醇分析（放射免疫测定和质谱法），表明了30多种类固醇特征的肾上腺皮质细胞的分泌。血清补充培养基中的总未结合的类固醇分泌为2.83微克/10（6）细胞/24小时，无血清培养基少约4倍。 NCI-H295细胞中妊娠代谢的主要途径是雄激素合成，形成了脱氢双足球酮，雄激素，睾丸激素和至少三个硫化雄激素，以及雌激素。另外，皮质醇，皮质酮，醛固酮和11个β-羟基甲苯胺的形成表明存在11β-羟化酶。因此，类固醇生成的多种途径由NCI-H295细胞表达，包括形成皮质类固醇，矿物皮质激素，雄激素和雌激素。我们的发现表明，在所有主要肾上腺皮质酶系统的NCI-H295细胞中存在，包括11个β-羟化酶，脱莫尔酶，21α-羟化酶，17个α-羟化酶，17个α-羟化酶，18-羟基酶，溶裂酶，溶酶体，硫基激酶和神拟酶。 NCI-H295细胞系应证明在研究参与类固醇形成和分泌的调节，代谢途径和酶时具有价值。此外，它可以提供有关肾上腺皮质癌的生物学和治疗的见解。
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 steroid-secreting 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 micrograms/10(6) 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 11 beta-hydroxyandrostenidione indicated the presence of 11 beta-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 11 beta-hydroxylase, desmolase, 21 alpha-hydroxylase, 17 alpha-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.