title: Cytotoxic activity of sunitinib and everolimus in Caki-1 renal cancer cells is accompanied by modulations in the expression of apoptosis-related microRNA clusters and BCL2 family genes.
abstract: Sunitinib and everolimus are two of the antineoplastic agents indicated for the management of metastatic renal cancer. Although both of the above compounds were primarily designed as antiangiogenic factors, preclinical studies claim that these drugs can also trigger apoptosis. Herein, we sought to evaluate the cytotoxic activity of sunitinib and everolimus against renal cancer cells Caki-1 and moreover to assess their impact on the expression levels of three BCL2 family members and three apoptosis-related microRNA clusters upon incubation with the drugs or following recovery from treatment. The cytotoxic effect of sunitinib and everolimus on Caki-1 cells' viability was estimated by the MTT assay, while cleaved PARP, assayed via Western Blotting, served as a marker of programmed cell death. As for the expression levels of the BCL2 family members BCL2, BAX and BCL2L12 and those of the mature microRNAs of the miR-183/96/182, miR-143/145, and miR-15a/16 clusters, they were quantified via real-time PCR. Our results showed that both agents induced a time- and dose-dependent decrease in cell viability and promoted cleavage of PARP. In parallel, significant modulations were observed in the expression levels of miR-145, miR-15a, and miR-16 in case of sunitinib, whereas BCL2, BAX, miR-145 and miR-15a expression was strongly affected by everolimus. Overall, our data support the notion that sunitinib and everolimus are able to directly induce cell death in renal cancer cells and simultaneously affect the expression levels of their apoptosis-related microRNAs and BCL2 family members upon this process.
authors.affiliation: Department of Biochemistry and Molecular Biology, Faculty of Biology, University of Athens, Panepistimiopolis, Athens 15701, Greece.
authors.forename: Emmanuel I
authors.initials: EI
authors.lastname: Papadopoulos
authors.affiliation: Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Ontario M5S 1A8, Canada.
authors.forename: George M
authors.initials: GM
authors.lastname: Yousef
authors.affiliation: Department of Biochemistry and Molecular Biology, Faculty of Biology, University of Athens, Panepistimiopolis, Athens 15701, Greece. Electronic address: ascorilas@biol.uoa.gr.
authors.forename: Andreas
authors.initials: A
authors.lastname: Scorilas
mesh_terms: D000970:Antineoplastic Agents / Q000008:administration & dosage / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D045744:Cell Line, Tumor; D000068338:Everolimus / Q000008:administration & dosage / Q000494:pharmacology; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D007211:Indoles / Q000008:administration & dosage / Q000494:pharmacology; D007680:Kidney Neoplasms / Q000188:drug therapy*; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D005810:Multigene Family; D009124:Muscle Proteins / Q000235:genetics / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics / Q000378:metabolism*; D011758:Pyrroles / Q000008:administration & dosage / Q000494:pharmacology; D000077210:Sunitinib; D051028:bcl-2-Associated X Protein / Q000235:genetics / Q000378:metabolism*
chemical_list: D000970:Antineoplastic Agents; C494826:BAX protein, human; C000595707:BCL2 protein, human; C430100:BCL2L12 protein, human; D007211:Indoles; D035683:MicroRNAs; D009124:Muscle Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D011758:Pyrroles; D051028:bcl-2-Associated X Protein; D000068338:Everolimus; D000077210:Sunitinib
keywords: BCL2 family; Caki-1 renal cancer cell line; Everolimus; MicroRNAs; Real-time PCR; Sunitinib