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PDGF induced microRNA alterations in cancer cells.

Shao M, Rossi S, Chelladurai B, Shimizu M, Ntukogu O, Ivan M, Calin GA, Matei D - Nucleic Acids Res. (2011)

Bottom Line: Here, we show by using microRNA (miR) arrays that PDGFs regulate the expression and function of miRs in glioblastoma and ovarian cancer cells.We demonstrate that PDGF regulates expression of some of its known targets (e.g. cyclin D1) through miR alterations and identify the epidermal growth factor receptor (EGFR) as a new PDGF-BB target.We show that its expression and function are repressed by PDGF-induced miR-146b and that mir-146b and EGFR correlate inversely in human glioblastomas.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Indiana University School of Medicine, University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA.

ABSTRACT
Platelet derived growth factor (PDGF) regulates gene transcription by binding to specific receptors. PDGF plays a critical role in oncogenesis in brain and other tumors, regulates angiogenesis, and remodels the stroma in physiologic conditions. Here, we show by using microRNA (miR) arrays that PDGFs regulate the expression and function of miRs in glioblastoma and ovarian cancer cells. The two PDGF ligands AA and BB affect expression of several miRs in ligand-specific manner; the most robust changes consisting of let-7d repression by PDGF-AA and miR-146b induction by PDGF-BB. Induction of miR-146b by PDGF-BB is modulated via MAPK-dependent induction of c-fos. We demonstrate that PDGF regulates expression of some of its known targets (e.g. cyclin D1) through miR alterations and identify the epidermal growth factor receptor (EGFR) as a new PDGF-BB target. We show that its expression and function are repressed by PDGF-induced miR-146b and that mir-146b and EGFR correlate inversely in human glioblastomas. We propose that PDGF-regulated gene transcription involves alterations in non-coding RNAs and provide evidence for a miR-dependent feedback mechanism balancing growth factor receptor signaling in cancer cells.

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PDGF-AA down-regulates let-7d expression. (A) qPCR quantifies let-7d expression in C272/hTert/E7 cells in response to PDGF-AA, PDGF-BB, VEGF and EGF. RNU49 was used as control. (B) Western blotting for cyclin-D1 in C272/hTert/E7 cells treated with PDGF-AA. (C) Western blotting for cyclin D1 in U118MG cells transfected with let-7d precursor or control (left panel). Western blotting for cyclin D1 in U118MG cells transfected with LNA targeting let-7d or scrambled LNA (right panel). (D) Western blotting assessed cyclin D1 in C272/hTERT/E7 cells treated with PDGF-AA for 6 h after transfection of let-7d precursor or control (left panel). Densitometry quantifies cyclin D1 expression levels relative to GAPDH in independent experiments. Statistical significance is indicated by asterisks (NS, not significant). (E) BRDU assay quantifies cell proliferation of C272/hTert/E7 cells transfected with let-7d precursor and control and stimulated with PDGF-AA or vehicle Statistical significance is marked by asterisks.
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Figure 2: PDGF-AA down-regulates let-7d expression. (A) qPCR quantifies let-7d expression in C272/hTert/E7 cells in response to PDGF-AA, PDGF-BB, VEGF and EGF. RNU49 was used as control. (B) Western blotting for cyclin-D1 in C272/hTert/E7 cells treated with PDGF-AA. (C) Western blotting for cyclin D1 in U118MG cells transfected with let-7d precursor or control (left panel). Western blotting for cyclin D1 in U118MG cells transfected with LNA targeting let-7d or scrambled LNA (right panel). (D) Western blotting assessed cyclin D1 in C272/hTERT/E7 cells treated with PDGF-AA for 6 h after transfection of let-7d precursor or control (left panel). Densitometry quantifies cyclin D1 expression levels relative to GAPDH in independent experiments. Statistical significance is indicated by asterisks (NS, not significant). (E) BRDU assay quantifies cell proliferation of C272/hTert/E7 cells transfected with let-7d precursor and control and stimulated with PDGF-AA or vehicle Statistical significance is marked by asterisks.

Mentions: To test the specificity of the miR response to individual growth factors, let-7d expression was quantified in C272hTert/E7 cells treated with PDGF-AA, PDGF-BB, EGF and VEGF. A ∼40% decrease in Let-7d level was observed in response to PDGF-AA, with no measurable changes observed in PDGF-BB, EGF or VEGF-treated cells (Figure 2A).Figure 2.


PDGF induced microRNA alterations in cancer cells.

Shao M, Rossi S, Chelladurai B, Shimizu M, Ntukogu O, Ivan M, Calin GA, Matei D - Nucleic Acids Res. (2011)

PDGF-AA down-regulates let-7d expression. (A) qPCR quantifies let-7d expression in C272/hTert/E7 cells in response to PDGF-AA, PDGF-BB, VEGF and EGF. RNU49 was used as control. (B) Western blotting for cyclin-D1 in C272/hTert/E7 cells treated with PDGF-AA. (C) Western blotting for cyclin D1 in U118MG cells transfected with let-7d precursor or control (left panel). Western blotting for cyclin D1 in U118MG cells transfected with LNA targeting let-7d or scrambled LNA (right panel). (D) Western blotting assessed cyclin D1 in C272/hTERT/E7 cells treated with PDGF-AA for 6 h after transfection of let-7d precursor or control (left panel). Densitometry quantifies cyclin D1 expression levels relative to GAPDH in independent experiments. Statistical significance is indicated by asterisks (NS, not significant). (E) BRDU assay quantifies cell proliferation of C272/hTert/E7 cells transfected with let-7d precursor and control and stimulated with PDGF-AA or vehicle Statistical significance is marked by asterisks.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

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Show All Figures
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Figure 2: PDGF-AA down-regulates let-7d expression. (A) qPCR quantifies let-7d expression in C272/hTert/E7 cells in response to PDGF-AA, PDGF-BB, VEGF and EGF. RNU49 was used as control. (B) Western blotting for cyclin-D1 in C272/hTert/E7 cells treated with PDGF-AA. (C) Western blotting for cyclin D1 in U118MG cells transfected with let-7d precursor or control (left panel). Western blotting for cyclin D1 in U118MG cells transfected with LNA targeting let-7d or scrambled LNA (right panel). (D) Western blotting assessed cyclin D1 in C272/hTERT/E7 cells treated with PDGF-AA for 6 h after transfection of let-7d precursor or control (left panel). Densitometry quantifies cyclin D1 expression levels relative to GAPDH in independent experiments. Statistical significance is indicated by asterisks (NS, not significant). (E) BRDU assay quantifies cell proliferation of C272/hTert/E7 cells transfected with let-7d precursor and control and stimulated with PDGF-AA or vehicle Statistical significance is marked by asterisks.
Mentions: To test the specificity of the miR response to individual growth factors, let-7d expression was quantified in C272hTert/E7 cells treated with PDGF-AA, PDGF-BB, EGF and VEGF. A ∼40% decrease in Let-7d level was observed in response to PDGF-AA, with no measurable changes observed in PDGF-BB, EGF or VEGF-treated cells (Figure 2A).Figure 2.

Bottom Line: Here, we show by using microRNA (miR) arrays that PDGFs regulate the expression and function of miRs in glioblastoma and ovarian cancer cells.We demonstrate that PDGF regulates expression of some of its known targets (e.g. cyclin D1) through miR alterations and identify the epidermal growth factor receptor (EGFR) as a new PDGF-BB target.We show that its expression and function are repressed by PDGF-induced miR-146b and that mir-146b and EGFR correlate inversely in human glioblastomas.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Indiana University School of Medicine, University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA.

ABSTRACT
Platelet derived growth factor (PDGF) regulates gene transcription by binding to specific receptors. PDGF plays a critical role in oncogenesis in brain and other tumors, regulates angiogenesis, and remodels the stroma in physiologic conditions. Here, we show by using microRNA (miR) arrays that PDGFs regulate the expression and function of miRs in glioblastoma and ovarian cancer cells. The two PDGF ligands AA and BB affect expression of several miRs in ligand-specific manner; the most robust changes consisting of let-7d repression by PDGF-AA and miR-146b induction by PDGF-BB. Induction of miR-146b by PDGF-BB is modulated via MAPK-dependent induction of c-fos. We demonstrate that PDGF regulates expression of some of its known targets (e.g. cyclin D1) through miR alterations and identify the epidermal growth factor receptor (EGFR) as a new PDGF-BB target. We show that its expression and function are repressed by PDGF-induced miR-146b and that mir-146b and EGFR correlate inversely in human glioblastomas. We propose that PDGF-regulated gene transcription involves alterations in non-coding RNAs and provide evidence for a miR-dependent feedback mechanism balancing growth factor receptor signaling in cancer cells.

Show MeSH
Related in: MedlinePlus