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Series GSE58199 Query DataSets for GSE58199
Status Public on Apr 19, 2016
Title Sema6A and Mical1 control cell growth and survival of BRAFV600E human melanomas
Organism Homo sapiens
Experiment type Expression profiling by array
Summary Therapeutic targeting of BRAFV600Eand of MEK has shown a significant impact on progression-free and overall survival in advanced melanoma, but only a fraction of patients benefit from these treatments, suggesting that additional signaling pathways involved in melanoma growth/survival need to be identified. To this end, we used whole genome microarray analysis to identify differentially expressed genes in a set of neoplastic clones, isolated from a single melanoma metastasis, and characterized by mututally exclusive expression of BRAFV600E or NRASQ61R. By this approach we identified two genes, SEMA6A and Mical-1 belonging to the semaphorin-plexin signaling pathway and higly expressed, at mRNA and protein level, in BRAF-mutant neoplastic clones. Real-time PCR, Western blot analysis and immunohistochemistry confirmed the preferential expression of SEMA-6A and Mical-1 in BRAFV600E neoplastic cells from melanoma clones, primary and metastatic cell lines and tissue sections from melanoma lesions. SEMA6A depletion, by specific RNA-interference experiments, led to cytoskeletal remodeling, loss of stress fibers, generation of actin-rich protrusion, and cell death, whereas SEMA6A overexpression, in NRASQ61R clones, promoted invasiveness. Mical-1 depletion, by siRNA, in BRAFV600E melanomas, did not alter the actin cytoskeleton organization but caused a strong NDR phosphorylation and NDR-dependent apoptosis. Overall, these results suggest that the SEMA and MICAL pathways contribute to promote survival of BRAFV600E melanomas.
 
Overall design The human melanoma cell lines Me 4405, Me26635, Me665/2, Me10538; clones 2/33; 2/21; 2/4; 2/17; 2/14; 2/59 were established in our laboratory from a surgical specimens. Cells were routinely maintained in RPMI medium 1640 supplemented with 10% FBS and 2 mM glutamine. Cells were maintained at 37°C in a water-saturated atmosphere of 5% CO2 in air. 6x106 cells were seeded in 75 cm2 flasks; after 72h Me 4405 and Me26635 were untreated or treated with fotemustine at 300 μmol/L for 6 hours. Each treatment or combination was performed in triplicate. The cells were collected and RNA extracted.
 
Contributor(s) De Cecco L, Anichini A
Citation(s) 25576923
Submission date Jun 03, 2014
Last update date Aug 16, 2018
Contact name Loris De Cecco
E-mail(s) loris.dececco@istitutotumori.mi.it
Organization name IRCSS Istituto Nazionale Tumori
Street address via Venezian 1
City Milan
ZIP/Postal code 20133
Country Italy
 
Platforms (1)
GPL6947 Illumina HumanHT-12 V3.0 expression beadchip
Samples (36)
GSM1403049 Me4405 6h (4775495006_A)
GSM1403050 Me4405 6h (4775495006_B)
GSM1403051 Me4405 6h (4775495006_C)
Relations
BioProject PRJNA251555

Download family Format
SOFT formatted family file(s) SOFTHelp
MINiML formatted family file(s) MINiMLHelp
Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE58199_RAW.tar 6.2 Mb (http)(custom) TAR
GSE58199_non-normalized_data.txt.gz 11.2 Mb (ftp)(http) TXT
Processed data included within Sample table

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