Transcriptional profile of Rous Sarcoma Virus transformed chicken embryo fibroblasts reveals new signaling targets of viral-src

Abstract

Transformation of chicken fibroblasts in vitro by Rous Sarcoma Virus represents a model of cancer in which a single oncogene, viral src, uniformly and rapidly transforms primary cells in culture. We experimentally surveyed the transcriptional program affected by Rous Sarcoma Virus (RSV) in primary culture of chicken embryo fibroblasts. As a control, we used cells infected with non-transforming RSV mutant td106, in which the src gene was deleted. Using Affymetrix GeneChip Chicken Genome Arrays, we report 811 genes that were modulated more than 2.5 fold in the virus transformed cells. Among these, 409 genes were induced and 402 genes were repressed by viral src. From the repertoire of modulated genes, we selected 20 genes that were robustly changed. We then validated and quantified the transcriptional changes of most of the 20 selected genes by real-time PCR. The set of strongly induced genes contains vasoactive intestinal polypeptide, MAP kinase phosphatase 2 and follistatin, among others. The set of strongly repressed genes contains TGF beta 3, TGF beta-induced gene, and deiodinase. The function of several robustly modulated genes sheds new light on the molecular mechanism of oncogenic transformation.

From Rous sarcoma virus to plasminogen activator, src oncogene and cancer management

“…In fact, our study showed that the PLAU transcript was upregulated 196 ( þ /À 69)-fold, using Affymetrix Gene Chip arrays. We confirmed this result by a more precise measurement using real-time PCR tests on two independently isolated RNA samples as 42 ( þ /À11)-fold upregulation (Masker et al, 2007). Independently from our group, the laboratory of Pierre-Andre Bedard also documented that the PLAU transcript was the most robustly upregulated gene in RSV-transformed CEFs, being more than 44-fold increased over the level detected in non-transformed controls (Maslikowski et al, 2010).…”

“…This work was reported by the laboratory of Edward Reich at The Rockefeller University in New York City, and later confirmed by numerous publications from various laboratories (Sudol, 1985;Bell et al, 1990;Leslie et al, 1990). More than three decades later, with the completion of the sequencing of mouse and chicken genomes and the subsequent availability of Affymetrix GeneChip genome arrays (Affymetrix, Santa Clara, CA, USA), several laboratories, including our team, have surveyed the transcriptional program affected by the RSV transformation (Masker et al, 2007;Maslikowski et al, 2010). We, and others, have confirmed the transcriptional induction of the PLAU gene and showed that it is the most highly upregulated transcript (Masker et al, 2007;Maslikowski et al, 2010).…”

“…As expected, the PLAU gene was shown to be induced by RSV at the level of transcription , and two recent reports with Affymetrix Gene chip analyses showed that in wt RSV Schmidt-Ruppinstrain-group-A (RSV-SR-A)-transformed chick embryo fibroblasts (CEFs), PLAU transcript was on the very top of the list of upregulated genes, being at least 40-fold elevated over the level found in cells infected with the non-transforming, control viruses of RSV-SR-A strain (Masker et al, 2007;Maslikowski et al, 2010). In fact, our study showed that the PLAU transcript was upregulated 196 ( þ /À 69)-fold, using Affymetrix Gene Chip arrays.…”

“…More than three decades later, with the completion of the sequencing of mouse and chicken genomes and the subsequent availability of Affymetrix GeneChip genome arrays (Affymetrix, Santa Clara, CA, USA), several laboratories, including our team, have surveyed the transcriptional program affected by the RSV transformation (Masker et al, 2007;Maslikowski et al, 2010). We, and others, have confirmed the transcriptional induction of the PLAU gene and showed that it is the most highly upregulated transcript (Masker et al, 2007;Maslikowski et al, 2010). Now, we have decided to look at these data more closely, focusing on unique features and variations in the PLAU gene induction among different experimental models.…”

Different responses in transformation of MDCK cells in 2D and 3D culture by v-Src as revealed by microarray techniques, RT-PCR and functional assays

“…They identified significant overexpression of DNA methyltransferase 1 in v-Src-transfected fibroblasts, whereas several tumor suppressor genes were downregulated by v-Src, but could be reexpressed after treatment with a demethylating agent. A similar analysis was carried out by Masker et al 42 using chicken embryo fibroblasts transfected with Rous Sarcoma virus. They observed 811 genes modulated by v-Src.…”