Elucidating the Role of the Maternal Embryonic Leucine Zipper Kinase in Adrenocortical Carcinoma

• Published in: Endocrinology (Philadelphia) Vol. 159; no. 7; pp. 2532 - 2544
• Main Authors: Kiseljak-Vassiliades, Katja, Zhang, Yu, Kar, Adwitiya, Razzaghi, Raud, Xu, Mei, Gowan, Katherine, Raeburn, Christopher D, Albuja-Cruz, Maria, Jones, Kenneth L, Somerset, Hilary, Fishbein, Lauren, Leong, Stephen, Wierman, Margaret E
• Format: Journal Article
• Language: English
• Published: Washington, DC Endocrine Society 01-07-2018; Oxford University Press
• Subjects: Adrenocortical Carcinoma - genetics; Adrenocortical Carcinoma - metabolism; Apoptosis; Apoptosis - drug effects; Apoptosis - genetics; Biotechnology; Cancer; Caspase; Caspase-3; Cell Cycle Proteins - genetics; Cell Cycle Proteins - metabolism; Cell division; Cell growth; Cell Line, Tumor; Cell proliferation; Cell Proliferation - drug effects; Cell Proliferation - genetics; Cell survival; Colonies; Dosage; Embryos; Endocrinology; Gene expression; Health aspects; Humans; Hypoxia; Immunoblotting; Immunohistochemistry; Kinases; Lamin Type B - genetics; Lamin Type B - metabolism; Leucine; Leucine zipper; Leucine zipper proteins; Naphthyridines - pharmacology; Neuroendocrine tumors; NIMA-Related Kinases - genetics; NIMA-Related Kinases - metabolism; Polo-Like Kinase 1; Protein Serine-Threonine Kinases - antagonists & inhibitors; Protein Serine-Threonine Kinases - genetics; Protein Serine-Threonine Kinases - metabolism; Proto-Oncogene Proteins - genetics; Proto-Oncogene Proteins - metabolism; Real-Time Polymerase Chain Reaction; Survival; Tumors; United States
• ISSN: 1945-7170; 0013-7227; 1945-7170
• DOI: 10.1210/en.2018-00310

Abstract
Adrenocortical carcinoma (ACC) is an aggressive cancer with a 5-year survival rate <35%. Mortality remains high due to lack of targeted therapies. Using bioinformatic analyses, we identified maternal embryonic leucine zipper kinase (MELK) as 4.1-fold overexpressed in ACC compared with normal adrenal samples. High MELK expression in human tumors correlated with shorter survival and with increased expression of genes involved in cell division and growth. We investigated the functional effects of MELK inhibition using newly developed ACC cell lines with variable MELK expression, CU-ACC1 and CU-ACC2, compared with H295R cells. In vitro treatment with the MELK inhibitor, OTSSP167, resulted in a dose-dependent decrease in rates of cell proliferation, colony formation, and cell survival, with relative sensitivity of each ACC cell line based upon the level of MELK overexpression. To confirm a MELK-specific antitumorigenic effect, MELK was inhibited in H295R cells via multiple short hairpin RNAs. MELK silencing resulted in 1.9-fold decrease in proliferation, and 3- to 10-fold decrease in colony formation in soft agar and clonogenicity assays, respectively. In addition, although MELK silencing had no effect on survival in normoxia, exposure to a hypoxia resulted in a sixfold and eightfold increase in apoptosis as assessed by caspase-3 activation and TUNEL, respectively. Together these data suggest that MELK is a modulator of tumor cell growth and survival in a hypoxic microenvironment in adrenal cancer cells and support future investigation of its role as a therapeutic kinase target in patients with ACC.
MELK is overexpressed in human adrenocortical carcinoma cells and drives proliferation, tumorigenesis, and survival in a hypoxic environment. MELK is a therapeutic target for patients with ACC.