Suppression subtractive hybridization profiles of radial growth phase and metastatic melanoma cell lines reveal novel potential targets.

BMC Cancer
Josane F Sousa, Enilza M Espreafico

Abstract

Melanoma progression occurs through three major stages: radial growth phase (RGP), confined to the epidermis; vertical growth phase (VGP), when the tumor has invaded into the dermis; and metastasis. In this work, we used suppression subtractive hybridization (SSH) to investigate the molecular signature of melanoma progression, by comparing a group of metastatic cell lines with an RGP-like cell line showing characteristics of early neoplastic lesions including expression of the metastasis suppressor KISS1, lack of alphavbeta3-integrin and low levels of RHOC. Two subtracted cDNA collections were obtained, one (RGP library) by subtracting the RGP cell line (WM1552C) cDNA from a cDNA pool from four metastatic cell lines (WM9, WM852, 1205Lu and WM1617), and the other (Met library) by the reverse subtraction. Clones were sequenced and annotated, and expression validation was done by Northern blot and RT-PCR. Gene Ontology annotation and searches in large-scale melanoma expression studies were done for the genes identified. We identified 367 clones from the RGP library and 386 from the Met library, of which 351 and 368, respectively, match human mRNA sequences, representing 288 and 217 annotated genes. We confirmed the differential ex...Continue Reading

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Citations

May 5, 2012·Modern Pathology : an Official Journal of the United States and Canadian Academy of Pathology, Inc·Namrata SetiaMeera Mahalingam
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Datasets Mentioned

BETA
ES315683

Methods Mentioned

BETA
PCR
electrophoresis
gel filtration
GTPase
flow cytometry

Software Mentioned

BLAT
GOTM
SAM ( Significance Analysis of Microarrays )
CLUSTER
Gene Ontology Tree Machine
WebGestalt
JAVATREEVIEW
SAMTERS
SAMSTER
Java TreeView

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