Identification of Src transformation fingerprint in human colon cancer

Malek, Renae L.; Irby, Rosalyn; Guo, Qingbin; Lee, Kerry; Wong, Sylvia; He, Mei; Tsai, Jennifer; Frank, Bryan; Liu, Edison T.; Quackenbush, John; Jove, Richard; Yeatman, Timothy J.; Lee, Norman H.

doi:10.1038/sj.onc.1205900

Cited by 72 publications

(63 citation statements)

References 51 publications

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“…The cells expressing Y527F Src showed prominent overall tyrosine phosphorylation compared with the cells which were not expressing active Src, as detected with an anti-(pY) antibody (data not shown). Introduction of active Src construct led to the upregulation of ezrin in the MEFs add-back clones (compare lane 2 in Figure 1a with lanes 4-6), in line with a previous fingerprint analysis of Src-regulated changes in gene expression (Malek et al, 2002).…”

Section: Characterization Of Mouse Embryonic Fibroblasts Subclonessupporting

confidence: 63%

Ezrin is key regulator of Src-induced malignant phenotype in three-dimensional environment

et al. 2011

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The oncogenic tyrosine kinase Src has a role in cancer development, especially by promoting invasive and metastatic behavior. It is, however, unclear which of the Srcregulated signaling cascades induce malignant phenotype in three-dimensional environment. One of Src substrates is ezrin, a cytoskeletal organiser and regulator of signal transduction. Ezrin expression correlates with poor outcome of diverse cancers and is essential in experimental metastatic osteosarcoma. We reconstituted genetically ezrin-deficient cells with wild-type (WT) or phosphorylation-deficient Y477F ezrin together with constitutively active Src. In two-dimensional cultures, Src induced malignant features regardless of the presence or absence of ezrin. In contrast, only WT ezrin-expressing cells grew efficiently in soft agar or in suspension. In Matrigel, only WT ezrin significantly promoted growth and invasion, and was targeted to specific regions on the plasma membrane. WT and Y477F ezrin-expressing cells showed marked differences only when growing or scattering in threedimensional matrix. Additional experiments showed that Y477-phosphorylated ezrin is also needed for the growth of Src-transformed epithelial cells in three-dimensional matrix. Cells lacking functional ezrin had reduced cyclin D levels and fewer cells in G2 þ S phase, possibly as a consequence of abnormal mTOR signaling, as ezrin Y477F cells showed lower expression of phosphorylated intermediates downstream of mTOR than WT cells. We conclude that the pathways activated by Src depend on the type of environment and that ezrin is a crucial element of Src-induced malignant features in cells growing inside three-dimensional environment.

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Section: Characterization Of Mouse Embryonic Fibroblasts Subclonessupporting

confidence: 63%

Ezrin is key regulator of Src-induced malignant phenotype in three-dimensional environment

et al. 2011

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“…(In an earlier experiment of a quite different design, Malek et al found a concordance between mRNAs that were present in human tumors and in Src transformed rat cells, but they did not report directly the changes (if any) between nontransformed rat 3Y1 cells and v-Src transformed cells.) (Malek et al, 2002) In contrast with the results, after 24 h of oncogene expression in our experiments, extensive changes were detected by gene array analysis after 15 days of continued expression. Armed with the prior knowledge (Bromberg et al, 1999;Shen et al, 2001) of some mRNAs that should have been increased after STAT3 activation, plus knowledge of mRNAs that were increased after 15 days of expression, we returned to test by RT-PCR for the presence of a set of such mRNAs early after expression of STAT3-C, v-Src or Ha-Ras.…”

Section: Discussioncontrasting

confidence: 51%

Transformation fingerprint: induced STAT3-C, v-Src and Ha-Ras cause small initial changes but similar established profiles in mRNA

et al. 2004

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“…In order to validate microarray and gene knockdown experiments with siRNA, real-time PCR (RT-PCR) was performed as described previously (Malek et al, 2002). Total RNA from cell lines were reversed transcribed using random primers as per the manufacturer's protocol.…”

Section: Quantitative Real-time Pcrmentioning

confidence: 99%

Autocrine activation of an osteopontin-CD44-Rac pathway enhances invasion and transformation by H-RasV12

Teramoto

Castellone

Malek³

et al. 2004

Oncogene

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Activated forms of Ras family members are prevalent in many cancers where Ras mutants transduce signals essential for transformation, angiogenesis, invasion and metastasis. As a cancer progression model, we used NIH3T3 cells to explore the mechanism of Ras-induced tumorigenesis. Ras family mutants H-RasV12 and Rit79L strongly induced foci formation, while Rho family mutants RhoA-QL, Rac1-QL and Cdc42-QL were less effective. A comparison of downstream transcriptional targets of Ras and Rho family members using a 26 383 element cDNA microarray revealed that the osteopontin (OPN) gene exhibited the best correlation between magnitude of gene expression change and level of foci formation (r ¼ 0.96, Po0.001). In association with H-RasV12-and Rit79L-mediated transformation, foci secreted OPN protein and upregulated the OPN receptor CD44, suggesting the novel initiation of an aberrant OPN-CD44-Rac autocrine pathway. In support of this were the following observations. First, RGD-deficient OPN protein-binding activity was present in H-RasV12-transformed cells but not in control cells, and binding activity was inhibited by the CD44 blocking antibody. Second, foci formation, cell invasion and Rac activity were induced by H-RasV12 and inhibited by the CD44 blocking antibody. Third, foci formation by H-RasV12 was substantially reduced by a short interfering RNA (siRNA) specifically targeting OPN expression for knockdown. Fourth, H-RasV12-mediated transformation was not blocked by the GRGDS peptide, suggesting that OPN effects were not mediated by the integrins. Lastly, OPN knockdown affected the downstream expression of 160 '2nd tier' genes, and at least a subset of these genes appears to be involved in transformation. Indeed, four genes were selected for knockdown, each resulting in a disruption of foci formation and/or invasion. These results underscore the role of aberrant autocrine signaling and transcriptional networking during tumorigenesis.

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Identification of Src transformation fingerprint in human colon cancer

Cited by 72 publications

References 51 publications

Ezrin is key regulator of Src-induced malignant phenotype in three-dimensional environment

Ezrin is key regulator of Src-induced malignant phenotype in three-dimensional environment

Transformation fingerprint: induced STAT3-C, v-Src and Ha-Ras cause small initial changes but similar established profiles in mRNA

Autocrine activation of an osteopontin-CD44-Rac pathway enhances invasion and transformation by H-RasV12

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