A cancer-specific transcriptional signature in human neoplasia
Francesco Nicassio, … , Ian Marc Bonapace, Pier Paolo Di Fiore
Francesco Nicassio, … , Ian Marc Bonapace, Pier Paolo Di Fiore
Published November 1, 2005
Citation Information: J Clin Invest. 2005;115(11):3015-3025. https://doi.org/10.1172/JCI24862.
View: Text | PDF
Research Article Oncology

A cancer-specific transcriptional signature in human neoplasia

Abstract

The molecular anatomy of cancer cells is being explored through unbiased approaches aimed at the identification of cancer-specific transcriptional signatures. An alternative biased approach is exploitation of molecular tools capable of inducing cellular transformation. Transcriptional signatures thus identified can be readily validated in real cancers and more easily reverse-engineered into signaling pathways, given preexisting molecular knowledge. We exploited the ability of the adenovirus early region 1 A protein (E1A) oncogene to force the reentry into the cell cycle of terminally differentiated cells in order to identify and characterize genes whose expression is upregulated in this process. A subset of these genes was activated through a retinoblastoma protein/E2 viral promoter required factor–independent (pRb/E2F-independent) mechanism and was overexpressed in a fraction of human cancers. Furthermore, this overexpression correlated with tumor progression in colon cancer, and 2 of these genes predicted unfavorable prognosis in breast cancer. A proof of principle biological validation was performed on one of the genes of the signature, skeletal muscle cell reentry-induced (SKIN) gene, a previously undescribed gene. SKIN was found overexpressed in some primary tumors and tumor cell lines and was amplified in a fraction of colon adenocarcinomas. Furthermore, knockdown of SKIN caused selective growth suppression in overexpressing tumor cell lines but not in tumor lines expressing physiological levels of the transcript. Thus, SKIN is a candidate oncogene in human cancer.

Authors

Francesco Nicassio, Fabrizio Bianchi, Maria Capra, Manuela Vecchi, Stefano Confalonieri, Marco Bianchi, Deborah Pajalunga, Marco Crescenzi, Ian Marc Bonapace, Pier Paolo Di Fiore

×

Figure 7

Options: View larger image (or click on image) Download as PowerPoint
Functional ablation of SKIN in tumor cell lines. Six different tumor cel...
Functional ablation of SKIN in tumor cell lines. Six different tumor cell lines (as indicated) were treated with SKIN-specific siRNA (open circles in A; RNAi in B and C) or a control scrambled oligo (filled triangles in A; scr. in B and C) or were mock treated (filled squares in A; mock in B and C). Twenty-four hours after treatment, cells were replated to measure cell growth (A) or analyzed for SKIN transcript levels by Q–RT-PCR (B). (A) Cells, replated in standard growth medium, were counted at the indicated time points. Data are expressed relative to the number of cells present in the plate 24 hours after replating (assumed as 1). (B) Q–RT-PCR data are expressed relative to those detected in growing MCF10A cells, to allow for comparison among cell lines. The mRNA levels of 2 double-stranded RNA-activated protein kinase–induced (PKR-induced) genes (STAT1 and interferon-induced transmembrane protein-IFITM1) were also analyzed to exclude nonspecific effects driven by SKIN-siRNA procedures and are reported on in Supplemental Figure 6. (C) In the case of DLD1 and HT-29 cells, levels of SKIN were also measured by Western blot with an anti-SKIN antibody. Results in A, B, and C were also replicated with a second SKIN-specific siRNA (oligo 2; see Supplemental Methods) with comparable results (not shown).