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035 a| (Sirsi) o1021186446;

035 a| (OCoLC)1021186446;

049 a| EREE;

100 1 a| Clark, Alexander G.,e| author.?| UNAUTHORIZED;

245 10 a| TGF-[beta] signaling, via Smad3, mediates Notch pathway-induced stemness and epithelial to mesenchymal transition in colon cancer cells /c| Alexander G. Clark.;

264 1 a| [Greenville, N.C.] :b| [East Carolina University],c| 2017.;

300 a| 91 pages :b| illustrations (some color).;

336 a| textb| txt2| rdacontent;

337 a| computerb| c2| rdamedia;

338 a| online resourceb| cr2| rdacarrier;

347 a| text fileb| PDFc| 1.832 MB2| rda;

490 1 a| ECU Brody School of Medicine thesis;

538 a| System requirements: Adobe Reader.;

538 a| Mode of access: World Wide Web.;

502 b| M.S.c| East Carolina Universityd| 2017.;

500 a| Presented to the faculty of the Department of Biomedical Science;

500 a| Advisor: George Sigounas;

500 a| Title from PDF t.p. (viewed February 28, 2018).;

520 3 a| Late stage colorectal cancer (CRC) remains a challenging disease to treat due to several factors including stemness and epithelial to mesenchymal transition (EMT). Dysfunctional signaling pathways in CRC contribute to these phenomena, including the Notch and Transforming Growth Factor-Beta (TGF-[beta]) pathways. These pathways integrate external signals by cross-talking with one another to fine-tune cellular responses. We previously found that cells expressing constitutively active Notch1 also had increased expression of Smad3, an important member of the TGF-[beta] signaling pathway. Therefore, we hypothesized that the TGF-[beta] pathway, via Smad3, mediates the Notch-induced stemness and EMT observed in these cells. To test our hypothesis, we used the human colorectal carcinoma cell line HCT-116 and derivative cell lines GFP-v (a control cell line transfected with a retrovirus containing the green fluorescent protein), and ICN1 (a cell line transfected with a retrovirus containing both the green fluorescent protein and constitutively active intracytoplasmic Notch1). These cells were treated with different combinations of TGF-[beta]1 (a TGF-[beta] receptor ligand), DAPT (a [gamma]-secretase inhibitor), or SIS3 (a novel Smad3 inhibitor). Western blot procedures and statistical analysis were performed to determine the cross-talk between the Notch and TGF-[beta] pathways and to assess the effects of Smad3 stimulation and inhibition on Notch and its downstream targets. The role of Smad3 on colosphere formation was also determined using the aforementioned cell lines. Smad3 inhibition induced a decrease in Notch1 and Notch3 receptor expression. Additionally, SIS3 effectively inhibited key stemness and EMT markers such as CD44, Slug, Snail, and Hes1. Colosphere forming ability was also considerably reduced in cells treated with SIS3. These results indicate a key role of TGF-[beta] signaling in Notch1-induced tumorigenesis, and they also suggest a potential use for Smad3 inhibitors in combination with Notch1 inhibitors that are already in use for CRC treatments.;

504 a| Includes bibliographical references.;

650 2 a| Colonic Neoplasms0| (DNLM)D003110=| ^A920109;

650 2 a| Epithelial-Mesenchymal Transition0| (DNLM)D058750=| ^A1280275;

650 2 a| Smad3 Protein0| (DNLM)D051900=| ^A1357120;

650 2 a| Signal Transduction0| (DNLM)D015398=| ^A925743;

650 2 a| Cell Count0| (DNLM)D002452=| ^A1357053;

653 a| Notch signaling;

700 1 a| Sigounas, George,e| degree supervisor.?| UNAUTHORIZED;

710 2 a| Brody School of Medicine.b| Office of Research and Graduate Studies.b| Biomedical Sciences.?| UNAUTHORIZED;

830 0 a| ECU Brody School of Medicine thesis.=| ^A964744;

856 40 z| Access via ScholarShipu| http://hdl.handle.net/10342/6464;

994 a| C0b| ERE;

096 a| WI 529;

596 a| 1 4;

998 a| 4814940;

999 a| CLICK ON WEB ADDRESSw| ASISc| 1i| 4814940-1001l| JNETm| JOYNERr| Ys| Yt| JNE3ETDu| 2/1/2018x| ETDz| JERESOURCE;

999 a| CLICK ON WEB ADDRESSw| ASISc| 1i| 4814940-2001l| HSLELECm| HSLr| Ys| Yt| HEETDu| 2/1/2018x| ETDz| HERESOURCE;

TGF-[beta] signaling, via Smad3, mediates Notch pathway-induced stemness and epithelial to mesenchymal transition in colon cancer cells / Alexander G. Clark.

Author/creator	Clark, Alexander G. author.
Other author	Sigounas, George, degree supervisor.
Other author	Brody School of Medicine. Office of Research and Graduate Studies. Biomedical Sciences.
Format	Theses and dissertations
Publication	[Greenville, N.C.] : [East Carolina University], 2017.
Description	91 pages : illustrations (some color).
Supplemental Content	Access via ScholarShip
Subjects	Colonic Neoplasms Epithelial-Mesenchymal Transition Smad3 Protein Signal Transduction Cell Count

Series	ECU Brody School of Medicine thesis ECU Brody School of Medicine thesis. ^A964744
Summary	Late stage colorectal cancer (CRC) remains a challenging disease to treat due to several factors including stemness and epithelial to mesenchymal transition (EMT). Dysfunctional signaling pathways in CRC contribute to these phenomena, including the Notch and Transforming Growth Factor-Beta (TGF-[beta]) pathways. These pathways integrate external signals by cross-talking with one another to fine-tune cellular responses. We previously found that cells expressing constitutively active Notch1 also had increased expression of Smad3, an important member of the TGF-[beta] signaling pathway. Therefore, we hypothesized that the TGF-[beta] pathway, via Smad3, mediates the Notch-induced stemness and EMT observed in these cells. To test our hypothesis, we used the human colorectal carcinoma cell line HCT-116 and derivative cell lines GFP-v (a control cell line transfected with a retrovirus containing the green fluorescent protein), and ICN1 (a cell line transfected with a retrovirus containing both the green fluorescent protein and constitutively active intracytoplasmic Notch1). These cells were treated with different combinations of TGF-[beta]1 (a TGF-[beta] receptor ligand), DAPT (a [gamma]-secretase inhibitor), or SIS3 (a novel Smad3 inhibitor). Western blot procedures and statistical analysis were performed to determine the cross-talk between the Notch and TGF-[beta] pathways and to assess the effects of Smad3 stimulation and inhibition on Notch and its downstream targets. The role of Smad3 on colosphere formation was also determined using the aforementioned cell lines. Smad3 inhibition induced a decrease in Notch1 and Notch3 receptor expression. Additionally, SIS3 effectively inhibited key stemness and EMT markers such as CD44, Slug, Snail, and Hes1. Colosphere forming ability was also considerably reduced in cells treated with SIS3. These results indicate a key role of TGF-[beta] signaling in Notch1-induced tumorigenesis, and they also suggest a potential use for Smad3 inhibitors in combination with Notch1 inhibitors that are already in use for CRC treatments.
General note	Presented to the faculty of the Department of Biomedical Science
General note	Advisor: George Sigounas
General note	Title from PDF t.p. (viewed February 28, 2018).
Dissertation note	M.S. East Carolina University 2017.
Bibliography note	Includes bibliographical references.
Technical details	System requirements: Adobe Reader.
Technical details	Mode of access: World Wide Web.

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TGF-[beta] signaling, via Smad3, mediates Notch pathway-induced stemness and epithelial to mesenchymal transition in colon cancer cells / Alexander G. Clark.

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