title: Effects of mitotane on gene expression in the adrenocortical cell line NCI-H295R: a microarray study.
abstract: OBJECTIVE
  The adrenolytic agent mitotane is widely used in the treatment of adrenocortical cancer; however, its mechanism of action is poorly elucidated. We have studied mitotane-induced mRNA expression changes in the NCI-H295R adrenocortical cancer cell line.
  
  
  METHODS
  Cell viability and hormone assays were used to select the optimal mitotane concentration effectively inhibiting hormone secretion without affecting cell viability. RNA isolated from cultures treated for 48 and 72 h was subjected to Agilent 4×44K microarray platforms. Microarray results were validated by quantitative reverse-transcription PCR.
  
  
  RESULTS
  Altogether, 117 significantly differentially expressed genes were detected at 48 h and 72 h (p < 0.05) in mitotane-treated samples relative to controls. Three significantly underexpressed genes involved in steroid hormone biosynthesis (HSD3B1, HSD3B2 and CYP21A2) and four significantly overexpressed genes (GDF15, ALDH1L2, TRIB3 and SERPINE2) have been validated.
  
  
  CONCLUSIONS
  Gene-expression changes might be involved in the adrenal action of mitotane and in the inhibition of hormone secretion.
authors.affiliation: 2nd Department of Medicine, Faculty of Medicine, Semmelweis University, Szentkirályi street 46, H-1088 Budapest, Hungary.
authors.forename: Adrienn
authors.initials: A
authors.lastname: Zsippai
authors.forename: Diana Rita
authors.initials: DR
authors.lastname: Szabó
authors.forename: Zsófia
authors.initials: Z
authors.lastname: Tömböl
authors.forename: Peter M
authors.initials: PM
authors.lastname: Szabó
authors.forename: Katalin
authors.initials: K
authors.lastname: Eder
authors.forename: Eva
authors.initials: E
authors.lastname: Pállinger
authors.forename: Rolf C
authors.initials: RC
authors.lastname: Gaillard
authors.forename: Attila
authors.initials: A
authors.lastname: Patócs
authors.forename: Sára
authors.initials: S
authors.lastname: Tóth
authors.forename: András
authors.initials: A
authors.lastname: Falus
authors.forename: Károly
authors.initials: K
authors.lastname: Rácz
authors.forename: Peter
authors.initials: P
authors.lastname: Igaz
mesh_terms: D000302:Adrenal Cortex / Q000187:drug effects; D000306:Adrenal Cortex Neoplasms / Q000188:drug therapy / Q000235:genetics*; D018931:Antineoplastic Agents, Hormonal / Q000494:pharmacology*; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects / Q000235:genetics; D015870:Gene Expression / Q000187:drug effects* / Q000235:genetics; D006728:Hormones / Q000235:genetics*; D006801:Humans; D046228:Microarray Analysis / Q000379:methods; D008939:Mitotane / Q000494:pharmacology*; D012333:RNA, Messenger / Q000235:genetics
chemical_list: D018931:Antineoplastic Agents, Hormonal; D006728:Hormones; D012333:RNA, Messenger; D008939:Mitotane