Synergistic response to oncogenic mutations defines gene class critical to cancer phenotype

Department of Biomedical Genetics, University of Rochester Medical Center, 601 Elmwood Avenue, Rochester, New York 14642, USA.
Nature (Impact Factor: 41.46). 07/2008; 453(7198):1112-6. DOI: 10.1038/nature06973
Source: PubMed


Understanding the molecular underpinnings of cancer is of critical importance to the development of targeted intervention strategies. Identification of such targets, however, is notoriously difficult and unpredictable. Malignant cell transformation requires the cooperation of a few oncogenic mutations that cause substantial reorganization of many cell features and induce complex changes in gene expression patterns. Genes critical to this multifaceted cellular phenotype have therefore only been identified after signalling pathway analysis or on an ad hoc basis. Our observations that cell transformation by cooperating oncogenic lesions depends on synergistic modulation of downstream signalling circuitry suggest that malignant transformation is a highly cooperative process, involving synergy at multiple levels of regulation, including gene expression. Here we show that a large proportion of genes controlled synergistically by loss-of-function p53 and Ras activation are critical to the malignant state of murine and human colon cells. Notably, 14 out of 24 'cooperation response genes' were found to contribute to tumour formation in gene perturbation experiments. In contrast, only 1 in 14 perturbations of the genes responding in a non-synergistic manner had a similar effect. Synergistic control of gene expression by oncogenic mutations thus emerges as an underlying key to malignancy, and provides an attractive rationale for identifying intervention targets in gene networks downstream of oncogenic gain- and loss-of-function mutations.

Download full-text


Available from: Lev Klebanov
  • Source
    • "Plasmids for retroviral Plac8 KD and shRNA-resistant Plac8 expression were described previously (McMurray et al., 2008). cDNAs for Rab7, Rab5a, and Atg12 were PCR cloned in pBabe-hygro containing an in-frame, N-terminal 3 3 Flag tag and sequence verified. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Mutations in p53 and RAS potently cooperate in oncogenic transformation, and correspondingly, these genetic alterations frequently coexist in pancreatic ductal adenocarcinoma (PDA) and other human cancers. Previously, we identified a set of genes synergistically activated by combined RAS and p53 mutations as frequent downstream mediators of tumorigenesis. Here, we show that the synergistically activated gene Plac8 is critical for pancreatic cancer growth. Silencing of Plac8 in cell lines suppresses tumor formation by blocking autophagy, a process essential for maintaining metabolic homeostasis in PDA, and genetic inactivation in an engineered mouse model inhibits PDA progression. We show that Plac8 is a critical regulator of the autophagic machinery, localizing to the lysosomal compartment and facilitating lysosome-autophagosome fusion. Plac8 thus provides a mechanistic link between primary oncogenic mutations and the induction of autophagy, a central mechanism of metabolic reprogramming, during PDA progression.
    Full-text · Article · Apr 2014 · Cell Reports
  • Source
    • "The third dataset is a comparison between four cell types—YAMC cells, mutant-p53 YAMC cells, activated-Ras YAMC cells and p53/Ras double mutant YAMC cells. Three replicates were performed for the untransformed YAMC cells, and four replicates were performed for each of the other cell types (McMurray et al., 2008). "
    [Show abstract] [Hide abstract]
    ABSTRACT: Quantitative real-time PCR (qPCR) is one of the most widely used methods to measure gene expression. Despite extensive research in qPCR laboratory protocols, normalization, and statistical analysis, little attention has been given to qPCR non-detects - those reactions failing to produce a minimum amount of signal. We show that the common methods of handling qPCR non-detects lead to biased inference. Furthermore, we show that non-detects do not represent data missing completely at random and likely represent missing data occurring not at random. We propose a model of the missing data mechanism and develop a method to directly model non-detects as missing data. Finally, we show that our approach results in a sizeable reduction in bias when estimating both absolute and differential gene expression. The proposed algorithm is implemented in the R package, nondetects. This package also contains the raw data for the three example data sets used in this manuscript. The package is freely available at and as part of the Bioconductor project.
    Full-text · Article · Apr 2014 · Bioinformatics
  • Source
    • "Therefore, we tested whether ncNRFR controls let-7 function. To do this, we used conditionally immortalized YAMC cells, which are useful to assay for oncogenic transformation in vitro and in vivo [26] [33]. YAMC cells were transfected with puromycin-selectable lentivirus-containing ncNRFR or an empty control vector. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Recent progress has been made in the identification of protein-coding genes and miRNAs that are expressed in and alter the behavior of colonic epithelia. However, the role of long non-coding RNAs (lncRNAs) in colonic homeostasis is just beginning to be explored. By gene expression profiling of post-mitotic, differentiated tops and proliferative, progenitor-compartment bottoms of microdissected adult mouse colonic crypts, we identified several lncRNAs more highly expressed in crypt bottoms. One identified lncRNA, designated non-coding Nras functional RNA (ncNRFR), resides within the Nras locus but appears to be independent of the Nras coding transcript. Stable overexpression of ncNRFR in non-transformed, conditionally immortalized mouse colonocytes results in malignant transformation, as determined by growth in soft agar and formation of highly invasive tumors in nude mice. Moreover, ncNRFR appears to inhibit the function of the tumor suppressor let-7. These results suggest precise regulation of ncNRFR is necessary for proper cell growth in the colonic crypt, and its misregulation results in neoplastic transformation.
    Full-text · Article · Sep 2013 · Biochemical and Biophysical Research Communications
Show more