Abstract

Newcastle disease virus (NDV) is used as an antineoplastic agent in clinical tumor therapy. It has prompted much interest as an anticancer agent because it can replicate up to 10,000 times better in human cancer cells than in most normal cells. This study was carried out to determine the oncolytic potential of NDV strain AF2240 and V4-UPM on WEHI-3B leukemia cell line. Results from MTT cytotoxicity assay showed that the CD50 values for both strains were 2 and 8 HAU for AF2240 and V4-UPM, respectively. In addition, bromodeoxyuridine (BrdU) and trypan blue dye exclusion assays showed inhibition in cell proliferation after different periods. Increase in the cellular level of caspase-3 and detection of DNA laddering using agarose gel electrophoresis on treated cells with NDV confirmed that the mode of cell death was apoptosis. In addition, flow-cytometry analysis of cellular DNA content showed that the virus caused an increase in the sub-G1 region (apoptosis peaks). In conclusion, NDV strains AF2240 and V4-UPM caused cytolytic effects against WEHI-3B leukemic cell line.

Highlights

  • Leukemia is a malignant neoplasm, characterized by neoplastic proliferation of hematopoietic cells, namely their diffuse replacement of the normal bone marrow [1,2]

  • There was a significant increase in apoptosis (Sub-G1) induction with time (p < 0.05) of WEHI-3B cell treated with both Newcastle disease virus (NDV) strains compared to untreated cells

  • In this study we reported that both NDV strains stimulate DNA fragmentation characteristic of apoptosis in WEHI-3B cell line at 24, 48 and 72 h post-inoculation

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Summary

Introduction

Leukemia is a malignant neoplasm, characterized by neoplastic proliferation of hematopoietic cells, namely their diffuse replacement of the normal bone marrow [1,2]. NDV is one of the nonengineered oncolytic viruses, which has a long history as a broad-spectrum oncolytic agent that can destroy tumor cells and stimulate the immune system [5]. It has oncolytic activity that can destroy tumor cells and stimulate the immune system. The oncolytic effects of six Malaysian strains of NDV (AF2240, 01/C, Ijuk, S, F, and V4) have been studied on several tumor cell lines [5,7]. No studies have yet been made using NDV oncolytic activity on myelomoncytic leukemia. In this study the oncolytic effects of two Malaysian local NDV strains AF2240 and V4-UPM were tested in vitro against Mouse myelomoncytic leukemia cell line, WEHI-3B

Cytotolytic Effects of NDV on Normal Cells and Myelomonocytic Leukemia Cells
BrdU Cell Proliferation Assay
Trypan Blue Exclusion Assay
DNA Fragmentation Assay
Effects of NDV on Cell Cycle Distribution
Propagation and Purification of NDV Strains AF2240 and V4-UPM
MTT Cytotoxicity Assay
BrdU Proliferation Assay
Analysis of Cellular DNA Content Using Propidium Iodide
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