Livin/BIRC7 expression as malignancy marker in adrenocortical tumors

// Barbara Altieri 1, 2 , Silviu Sbiera 3 , Silvia Della Casa 2 , Isabel Weigand 1 , Vanessa Wild 3, 4 , Sonja Steinhauer 1 , Guido Fadda 5 , Arkadius Kocot 6 , Michaela Bekteshi 1 , Egle M. Mambretti 7 , Andreas Rosenwald 4 , Alfredo Pontecorvi 2 , Martin Fassnacht 1, 3 , Cristina L. Ronchi 1 1 Department of Internal Medicine I, Division of Endocrinology and Diabetes, University Hospital of Wuerzburg, Germany 2 Division of Endocrinology and Metabolic Diseases, Catholic University of the Sacred Heart, Rome, Italy 3 Comprehensive Cancer Center Mainfranken, University of Wuerzburg, Germany 4 Department of Pathology, University of Wuerzburg, Germany 5 Division of Anatomic Pathology and Histology, Catholic University of the Sacred Heart, Rome, Italy 6 Department of Urology, University Hospital of Wuerzburg, Germany 7 Department of Anesthesiology and Critical Care, University Hospital of Wuerzburg, Germany Correspondence to: Cristina L. Ronchi, email: Ronchi_C@ukw.de Keywords: livin, BIRC7, adrenocortical carcinoma, adrenal tumor, caspase-3 Received: May 27, 2016      Accepted: December 15, 2016      Published: December 21, 2016 ABSTRACT Livin/BIRC7 is a member of the inhibitors of apoptosis proteins family, which are involved in tumor development through the inhibition of caspases. Aim was to investigate the expression of livin and other members of its pathway in adrenocortical tumors and in the adrenocortical carcinoma (ACC) cell line NCI-H295R. The mRNA expression of livin , its isoforms α and β , XIAP , CASP3 and DIABLO was evaluated by qRT-PCR in 82 fresh-frozen adrenal tissues (34 ACC, 25 adenomas = ACA, 23 normal adrenal glands = NAG). Livin protein expression was assessed by immunohistochemistry in 270 paraffin-embedded tissues (192 ACC, 58 ACA, 20 NAG). Livin, CASP3 and cleaved caspase-3 were evaluated in NCI-H295R after induction of livin overexpression. Relative livin mRNA expression was significantly higher in ACC than in ACA and NAG (0.060 ± 0.116 vs 0.004 ± 0.014 and 0.002 ± 0.009, respectively, p < 0.01), being consistently higher in tumors than in adjacent NAG and isoform β more expressed than α . No significant differences in CASP3 , XIAP and DIABLO levels were found among these groups. In immunohistochemistry, livin was localized in both cytoplasm and nuclei. The ratio between cytoplasmic and nuclear staining was significantly higher in ACC (1.51 ± 0.66) than in ACA (0.80 ± 0.35) and NAG (0.88 ± 0.27; p < 0.0001). No significant correlations were observed between livin expression and histopathological parameters or clinical outcome. In NCI-H295R cells, the livin overexpression slightly reduced the activation of CASP3, but did not correlate with cell viability. In conclusion, livin is specifically over-expressed in ACC, suggesting that it might be involved in adrenocortical tumorigenesis and represent a new molecular marker of malignancy.