REGULATION OF EGFR SIGNALING NETWORK BY NOVEL MOLECULES FOR CANCER THERAPY
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Title |
REGULATION OF EGFR SIGNALING NETWORK BY NOVEL MOLECULES FOR CANCER THERAPY |
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Creator |
Bhowmik, Arijit
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Subject |
Drug Development/Diagnostics & Biotechnology
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Description |
The thesis work is divided into two main sections, the first one emphasize on the regulation of EGFR signaling in breast cancer by novel indole derivative and the second one on the delivery of nanoencapsulated therapeutic indole derivative to EGFR overexpressed brain tumors . In the first part of the work, by in vitro screening we have selected a novel synthetic indole derivative 2,2‟-diphenyl-3,3‟- diindolylmethane (DPDIM) as a potential anti- breast cancer agent. DPDIM induces apoptosis both in vitro in breast cancer cells MCF7, MDA-MB 231 and MDA-MB 468 and in vivo in 7,12-dimethylbenz[a]anthracene (DMBA) induced Sprague- Dawley (SD) rat mammary tumor. Our in vitro studies show that DPDIM exerts apoptotic effect by negatively regulating the activity of EGFR and its downstream molecules like STAT3, AKT and ERK1/2 which are involved in the proliferation and survival of these cancer cells. In silico predictions also suggest that DPDIM may bind to EGFR at its ATP binding site. DPDIM furthermore inhibits EGF induced increased cell viability. We have also shown decreased expression of pro-survival factor Bcl-XL as well as increase in the level of pro-apoptotic proteins like Bax, Bad, Bim in DPDIM treated cells in vitro and in vivo. Our results further indicate that the DPDIM induced apoptosis is mediated through mitochondrial apoptotic pathway involving the caspase-cascade. Altogether this research work suggests that DPDIM could be an effective therapeutic agent for breast cancer. In the second part, in search of more effective therapeutic options for EGFR pathway overexpressed glioma treatment we prepared a nanoparticle encapsulated indole derivative (3,3‟diindolylmethane or DIM), tagged with a peptide against Somatostatin receptor 2 (SSTR2) on the surface of the nanoparticles for effective vi glioma therapy. DIM was encapsulated by poly(lactic-co-glycolic acid). The encapsulation was confirmed by TEM, FTIR, DLS etc. The effect of nanoparticle encapsulated DIM was examined on C6 rat glioma cell line by MTT assay, FACS, fluroscence microscopy, immunoblotting etc. To achieve the goal of target based delivery, a peptide against SSTR2 (overexpressed in glioma) was designed, synthesised and validated its target specificity in silico (by docking and simulation study) and in vitro (by fluorescence microscopy, IP and FACS). After tagging it chemically with the surface of DIM containing nanoparticles, passaging it through blood brain barrier (BBB) was determined by both in vitro BBB model and in vivo rat intracranial glioma model. Evaluation of the transportation of peptide coated nanoparticle encapsulated DIM through the BBB showed prominent passaging of the compound that induces apoptosis of glioma cells. |
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Date |
2015
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Type |
Thesis
NonPeerReviewed |
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Format |
application/pdf
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Identifier |
http://www.eprints.iicb.res.in/2470/1/Thesis_Arijit_Bhowmik.pdf
Bhowmik, Arijit (2015) REGULATION OF EGFR SIGNALING NETWORK BY NOVEL MOLECULES FOR CANCER THERAPY. PhD thesis, Calcutta University. |
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Relation |
http://www.eprints.iicb.res.in/2470/
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