Local V4-UPM strain Newcastle virus-induced cell death of brain tumor cell lines

V4-UPM strain is a local heat resistant variant of the avirulent Australian V4 (QUE) strain which has higher thermostabilities of infectivity and haemagglutination than the original. The aims of this study were to analyze the cytolytic effect and mode of cell death induced by V4-UPM strain of NDV on...

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Main Author: Ibrahim, Rohaya
Format: Thesis
Language:English
English
Published: 2011
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Online Access:http://psasir.upm.edu.my/id/eprint/25768/1/IB%202011%2033R.pdf
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spelling my-upm-ir.257682022-01-26T05:07:53Z Local V4-UPM strain Newcastle virus-induced cell death of brain tumor cell lines 2011-08 Ibrahim, Rohaya V4-UPM strain is a local heat resistant variant of the avirulent Australian V4 (QUE) strain which has higher thermostabilities of infectivity and haemagglutination than the original. The aims of this study were to analyze the cytolytic effect and mode of cell death induced by V4-UPM strain of NDV on two human brain tumor cell lines, glioblastoma multiforme (DBTRG.05MG) and anaplastic astrocytoma cells (U87MG).In-vitro cytotoxic activity has been performed on both brain tumor cells via standard MTT assay. At 72 hours post-incubation, the IC50 values obtained were 30 haem agglutination (HA) unit/ml and 5 HA unit/ml for DBTRG.05MG and U87MG cells,respectively. In addition, V4-UPM strain inhibited the proliferation of both DBTRG.05MG and U87MG cells via the same method. Morphological changes of the treated cells also have been observed sequentially under the inverted light, fluorescence and electron microscopes. The virus-infected cells showed apoptotic features such as the cell shrinkage, cell blebbing and formation of apoptotic bodies under the inverted light microscope. The differential uptake of acridine orange and propidium iodide dyes demonstrated the occurrence of apoptosis, mainly by chromatin condensation and DNA fragmentation that were observed under fluorescence microscope. The next morphological observation was based on color and nuclear morphology where the nuclei of treated-cells possessed DNA fragmentation were stained dark brown by 3’,3’-diaminobenzidine. The distinct ultrastructural changes occurred in the nucleus, cellular organelles and plasma membrane structures were confirmed to be the apoptotic features via transmission electron microscopic (TEM) observation. The result of flow cytometric analyses based on annexin V-FITC that binds phosphatidylserine residue also suggested the onset of apoptosis for both groups. Meanwhile, the cell cycle was arrested at S phase followed by apoptosis. Based on these findings, it was confirmed that the V4-UPM strain induced apoptotic cell death on DBTRG.05MG and U87MG cells. Thus, this strain has the potential to be developed as an antitumor agent for future treatment particularly in grade-III brain tumor patients. Newcastle disease virus Cell death. Brain - Cancer 2011-08 Thesis http://psasir.upm.edu.my/id/eprint/25768/ http://psasir.upm.edu.my/id/eprint/25768/1/IB%202011%2033R.pdf application/pdf en public masters Universiti Putra Malaysia Newcastle disease virus Cell death. Brain - Cancer Institute of Bioscience English
institution Universiti Putra Malaysia
collection PSAS Institutional Repository
language English
English
topic Newcastle disease virus
Cell death.
Brain - Cancer
spellingShingle Newcastle disease virus
Cell death.
Brain - Cancer
Ibrahim, Rohaya
Local V4-UPM strain Newcastle virus-induced cell death of brain tumor cell lines
description V4-UPM strain is a local heat resistant variant of the avirulent Australian V4 (QUE) strain which has higher thermostabilities of infectivity and haemagglutination than the original. The aims of this study were to analyze the cytolytic effect and mode of cell death induced by V4-UPM strain of NDV on two human brain tumor cell lines, glioblastoma multiforme (DBTRG.05MG) and anaplastic astrocytoma cells (U87MG).In-vitro cytotoxic activity has been performed on both brain tumor cells via standard MTT assay. At 72 hours post-incubation, the IC50 values obtained were 30 haem agglutination (HA) unit/ml and 5 HA unit/ml for DBTRG.05MG and U87MG cells,respectively. In addition, V4-UPM strain inhibited the proliferation of both DBTRG.05MG and U87MG cells via the same method. Morphological changes of the treated cells also have been observed sequentially under the inverted light, fluorescence and electron microscopes. The virus-infected cells showed apoptotic features such as the cell shrinkage, cell blebbing and formation of apoptotic bodies under the inverted light microscope. The differential uptake of acridine orange and propidium iodide dyes demonstrated the occurrence of apoptosis, mainly by chromatin condensation and DNA fragmentation that were observed under fluorescence microscope. The next morphological observation was based on color and nuclear morphology where the nuclei of treated-cells possessed DNA fragmentation were stained dark brown by 3’,3’-diaminobenzidine. The distinct ultrastructural changes occurred in the nucleus, cellular organelles and plasma membrane structures were confirmed to be the apoptotic features via transmission electron microscopic (TEM) observation. The result of flow cytometric analyses based on annexin V-FITC that binds phosphatidylserine residue also suggested the onset of apoptosis for both groups. Meanwhile, the cell cycle was arrested at S phase followed by apoptosis. Based on these findings, it was confirmed that the V4-UPM strain induced apoptotic cell death on DBTRG.05MG and U87MG cells. Thus, this strain has the potential to be developed as an antitumor agent for future treatment particularly in grade-III brain tumor patients.
format Thesis
qualification_level Master's degree
author Ibrahim, Rohaya
author_facet Ibrahim, Rohaya
author_sort Ibrahim, Rohaya
title Local V4-UPM strain Newcastle virus-induced cell death of brain tumor cell lines
title_short Local V4-UPM strain Newcastle virus-induced cell death of brain tumor cell lines
title_full Local V4-UPM strain Newcastle virus-induced cell death of brain tumor cell lines
title_fullStr Local V4-UPM strain Newcastle virus-induced cell death of brain tumor cell lines
title_full_unstemmed Local V4-UPM strain Newcastle virus-induced cell death of brain tumor cell lines
title_sort local v4-upm strain newcastle virus-induced cell death of brain tumor cell lines
granting_institution Universiti Putra Malaysia
granting_department Institute of Bioscience
publishDate 2011
url http://psasir.upm.edu.my/id/eprint/25768/1/IB%202011%2033R.pdf
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