Vol.:(0123456789) 1 3 Stem Cell Reviews and Reports https://doi.org/10.1007/s12015-018-9804-2 Efect of Everolimus on Heterogenous Renal Cancer Cells Populations Including Renal Cancer Stem Cells Anna Kornakiewicz 1,2  · Anna M. Czarnecka 1  · Mohammed I. Khan 1  · Paweł Krasowski 1  · Anna V. Kotrys 3  · Cezary Szczylik 1,4 © Springer Science+Business Media, LLC, part of Springer Nature 2018 Abstract The aim of this study was to compare efect of everolimus on growth of diferent renal cell carcinoma (RCC) populations and develop experimental design to measure the early response of everolimus in clear cell RCC (ccRCC) cell lines includ- ing renal cancer stem cells. Efect of everolimus on RCC cell lines which include primary (786-0) and metastatic (ACHN) RCC cell lines as well as heterogenous populations of tumor cells of diferent histological RCC subtypes (clear cell RCC and papillary RCC) was measured when treated with everolimus in the range of 1–9 µM. Gene expression profling using microarray was performed to determine the early response to everolimus in ccRCC cell lines after optimizing concentration of drug. Gene Set Enrichment Analysis (GSEA) was done which mainly focused on basic genes related to mTOR, hormo- nal and metabolic pathways. Everolimus acts on RCC cells in a dose—dependent manner. In all examined cell lines IC50 dose was possible to calculate after the third day of treatment. In ccRCC lines (parental and stem cell) everolimus changes expression of mTOR complexes elements and elements of related pathways when treated with optimized doses of drug. Characteristic expression profle for ccRCC cells at an early exposure time to everolimus is to elucidate. Wevarie include some basic observations derived from data analysis in the context of mechanism of action of drug with a view to better understand biology of renal cancer cells. Keywords Everolimus · Renal cell carcinoma · Clear cell renal cell carcinoma · Renal cancer stem cells · Experimental design Abbreviations 4EBP1 Eukaryotic translation initiation factor 4E-binding protein 1 786-O[0] Primary tumor cell line ACHN Malignant pleural efusion of metastatic renal adenocarcinoma cell line ACTH Adrenocorticotropic hormone ASE Healthy kidney cells ccRCC-PCSC Clear cell renal cell carcinoma - parental cell line Electronic supplementary material The online version of this article (https://doi.org/10.1007/s12015-018-9804-2) contains supplementary material, which is available to authorized users. * Anna Kornakiewicz anna.kornakiewicz@gmail.com Anna M. Czarnecka aczarnecka@wim.mil.pl Mohammed I. Khan imrankhanbioinfo@gmail.com Paweł Krasowski pkrasowski91@gmail.com Anna V. Kotrys a.v.kotrys@gmail.com Cezary Szczylik cszczylik@wim.mil.pl 1 Molecular Oncology Laboratory, Department of Oncology, Military Institute of Medicine, ul. Szaserów 128, 04-141 Warsaw, Poland 2 Postgraduate School of Molecular Medicine, Warsaw Medical University, Żwirki i Wigury 61, 02-091 Warsaw, Poland 3 Institute of Biochemistry and Biophysics Polish Academy of Sciences, Laboratory of RNA Biology and Functional Genomics, Pawińskiego 5A, 02-106 Warsaw, Poland 4 Warsaw Medical University, Żwirki i Wigury 61, 02-091 Warsaw, Poland