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Published on: November 24, 2014
ONCOLYTIC ACTIVITY OF HUMAN ORTHOPNEUMOVIRUS IN CANCER CELL LINES
I M Aziz1, R Bhat1, M A Farrag1
1College of Science, Alfaisal University, Riyadh 11533, Saudi Arabia. vrg_ksu@yahoo.com
Abstract:
Oncolytic virotherapy is an emerging biotherapeutic platform for selectively infecting cancer cells and triggering apoptosis in a number of malignant cells due to robust viral replication. Studies related to the oncolytic activity of human orthopneumovirus (hOPV) are conflicting.
Aim:
This study was designed to elucidate the possible role of hOPV in the modulation of cell growth and apoptosis in cancer cell lines including human epidermoid carcinoma (HEp-2), lung epithelial cell line (A549), and breast cancer cell line (MCF-7).
Materials And Methods:
The oncolytic activity of hOPV on cancer cells was studied in vitro. The virus titers were determined by tissue culture infectious dose (TCID50/mL) in A549 cell. The cytotoxic effect of the virus on HEp-2, A549, and MCF-7 was determined using MTT and trypan blue dye exclusion test assays. hOPV in the infected cells was detected using real-time reverse transcription polymerase chain reaction (rRT-PCR) and indirect immunofluorescence (IIF) assays. The relative expression of apoptosis-related genes (CASP-3, -8, -9, Bax, Bcl-2, Bcl-XL, TP53, P21) during virus infection was estimated using rRT-PCR assay in comparison with the house-keeping gene (GAPDH).
Results:
hOPV infection inhibited the growth of HEp-2, A549, and MCF-7 cells in a dose-and time-dependent manner. At a multiplicity of infection (MOI) of 5, hOPV reduced the viability of A549 cells to about 16%, HEp-2 to 22%, and MCF-7 to 28% (p = 0.001), while no significant inhibitory effect was observed when cells were infected at MOI of 1 and 2. hOPV mRNA and antigens were detected in infected HEp-2, A549, and MCF-7 cells by RT-PCR and IIF. Upon hOPV infection, expression of CASP-3, -8, -9, as well as Bax, TP53, and p21 mRNA increased while expression of Bcl-2, Bcl-xL anti-apoptotic genes decreased. In hOPV-infected A549 cells, the fold increase of CASP-8 and CASP-9, Bax, TP53, and P21 expression exceeded significantly compared to that in HEp-2 or MCF-7 cells.
Conclusions:
Our results provide evidence that hOPV could be a potential candidate for oncolytic virotherapy.
Insights
Human orthopneumovirus (hOPV) shows potential as an oncolytic virotherapy agent by inhibiting cancer cell growth and inducing apoptosis. This study confirmed hOPV
Area of Science:
- Oncolytic Virotherapy
- Cancer Biology
- Virology
Background:
- Oncolytic virotherapy utilizes viruses to selectively target and destroy cancer cells.
- Previous studies on the oncolytic activity of human orthopneumovirus (hOPV) have yielded conflicting results.
- Cancer cell lines, including HEp-2, A549, and MCF-7, were used to investigate hOPV's effects.
Purpose of the Study:
- To elucidate the role of hOPV in modulating cancer cell growth and apoptosis.
- To evaluate the potential of hOPV as an oncolytic agent against specific cancer types.
Main Methods:
- In vitro assessment of hOPV's oncolytic activity on HEp-2, A549, and MCF-7 cell lines.
- Quantification of virus titers using TCID50/mL.
- Assessment of cell viability and cytotoxicity via MTT and trypan blue assays.
- Detection of hOPV presence using RT-PCR and indirect immunofluorescence.
- Analysis of apoptosis-related gene expression (CASP-3, -8, -9, Bax, Bcl-2, Bcl-XL, TP53, P21) via RT-PCR.
Main Results:
- hOPV significantly inhibited the growth of HEp-2, A549, and MCF-7 cells in a dose- and time-dependent manner.
- At a multiplicity of infection (MOI) of 5, hOPV reduced cancer cell viability, with A549 cells showing the most significant reduction (16%).
- hOPV infection upregulated pro-apoptotic genes (CASP-3, -8, -9, Bax, TP53, P21) and downregulated anti-apoptotic genes (Bcl-2, Bcl-xL).
- A549 cells exhibited a significantly higher induction of apoptosis-related gene expression compared to HEp-2 and MCF-7 cells upon hOPV infection.
Conclusions:
- hOPV demonstrates significant oncolytic activity against tested cancer cell lines.
- The virus effectively modulates apoptosis-related gene expression, favoring cell death.
- hOPV presents a promising candidate for the development of novel oncolytic virotherapies.
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