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Oncolytic activity of Sindbis virus in human oral squamous carcinoma cells.

Saito K, Uzawa K, Kasamatsu A, Shinozuka K, Sakuma K, Yamatoji M, Shiiba M, Shino Y, Shirasawa H, Tanzawa H - Br. J. Cancer (2009)

Bottom Line: Sindbis viral growth was not observed in NHOKs, whereas high SIN growth was observed in all OSCC cell lines, including HCS-4.The CPE was shown, by terminal deoxyribonucleotidyl transferase-mediated dUTP nick-end labelling assays, to be apoptotic cell death.Furthermore, quantitative RT-PCR of mRNA in HSC-3 and HSC-4 cells after SIN infection showed that activation of caspases, cytochrome c, and IkappaBalpha was associated with SIN-induced apoptosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Virology, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba 260-8670, Japan.

ABSTRACT

Background: Sindbis virus (SIN) infection causes no or only mild symptoms (fever, rash, and arthralgia) in humans. However, SIN has a strong cytopathic effect (CPE) on various cancer cells. This study focuses on the oncolytic activity of SIN AR399 on oral cancer cells compared with reovirus, a well-known oncolytic virus that targets cancer cells.

Methods: We analysed the cytotoxicity and growth of SIN in 13 oral squamous cell carcinoma (OSCC) cell lines (HSC-2, HSC-3, HSC-4, Ca9-22, H-1, Sa-3, KON, KOSC-2, OK-92, HO-1-N1, SCC-4, SAT, SKN-3) and normal human oral keratinocytes (NHOKs).

Results: Sindbis virus infection induced CPE in 12 OSCC cell lines at a low multiplicity of infection (MOI) of 0.01, but not in the OSCC cell line, HSC-4 or NHOKs. Sindbis viral growth was not observed in NHOKs, whereas high SIN growth was observed in all OSCC cell lines, including HCS-4. The cytotoxicity and growth of SIN was the same as reovirus at an MOI of 20 in 12 OSCC cell lines. The CPE was shown, by terminal deoxyribonucleotidyl transferase-mediated dUTP nick-end labelling assays, to be apoptotic cell death. Furthermore, quantitative RT-PCR of mRNA in HSC-3 and HSC-4 cells after SIN infection showed that activation of caspases, cytochrome c, and IkappaBalpha was associated with SIN-induced apoptosis.

Conclusion: As a replication-competent oncolytic virus, SIN may be a useful therapeutic modality for oral cancers.

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Viral growth (A and B) in cells infected with SIN. (A) Titre of virus grown in various cells (keratinocytes, HSC-2, HSC-3, SCC-4, SAT, OK-92, Ca9-22, SKN-3, KON, KOSC-2, Sa-3, H-1, HO-1-N1, and HSC-4) was measured 96 h after SIN infection at an MOI of 0.1. (B) The titre of virus in cells (keratinocytes, HSC-2, HSC-3, SCC-4, SAT, OK-92, Ca9-22, SKN-3, KON, KOSC-2, Sa-3, H-1, HO-1-N1, and HSC-4) was measured 96 h after reovirus infection at an MOI of 0.1.
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fig2: Viral growth (A and B) in cells infected with SIN. (A) Titre of virus grown in various cells (keratinocytes, HSC-2, HSC-3, SCC-4, SAT, OK-92, Ca9-22, SKN-3, KON, KOSC-2, Sa-3, H-1, HO-1-N1, and HSC-4) was measured 96 h after SIN infection at an MOI of 0.1. (B) The titre of virus in cells (keratinocytes, HSC-2, HSC-3, SCC-4, SAT, OK-92, Ca9-22, SKN-3, KON, KOSC-2, Sa-3, H-1, HO-1-N1, and HSC-4) was measured 96 h after reovirus infection at an MOI of 0.1.

Mentions: To examine whether the extent of cell cytotoxicity after SIN infection correlates with viral growth, viral titres in the cancer cell lines were measured 96 h after SIN or reovirus infection (0.1 MOI). High growth of SIN and reovirus was observed in all OSCC cell lines (Figure 2) whereas no viral growth was observed in normal keratinocytes, supporting the idea that the cytotoxicity caused by SIN depends on replication of the virus. However, there was little correlation between the viral titre and cytotoxicity caused by infection. For example, the SIN viral growth was high in HSC-4 cells but SIN infection caused little cytotoxicity in these cells. In contrast, viral growth was low in Sa-3 and H-1 cells but SIN caused high cytotoxicity in these cell lines.


Oncolytic activity of Sindbis virus in human oral squamous carcinoma cells.

Saito K, Uzawa K, Kasamatsu A, Shinozuka K, Sakuma K, Yamatoji M, Shiiba M, Shino Y, Shirasawa H, Tanzawa H - Br. J. Cancer (2009)

Viral growth (A and B) in cells infected with SIN. (A) Titre of virus grown in various cells (keratinocytes, HSC-2, HSC-3, SCC-4, SAT, OK-92, Ca9-22, SKN-3, KON, KOSC-2, Sa-3, H-1, HO-1-N1, and HSC-4) was measured 96 h after SIN infection at an MOI of 0.1. (B) The titre of virus in cells (keratinocytes, HSC-2, HSC-3, SCC-4, SAT, OK-92, Ca9-22, SKN-3, KON, KOSC-2, Sa-3, H-1, HO-1-N1, and HSC-4) was measured 96 h after reovirus infection at an MOI of 0.1.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2736826&req=5

fig2: Viral growth (A and B) in cells infected with SIN. (A) Titre of virus grown in various cells (keratinocytes, HSC-2, HSC-3, SCC-4, SAT, OK-92, Ca9-22, SKN-3, KON, KOSC-2, Sa-3, H-1, HO-1-N1, and HSC-4) was measured 96 h after SIN infection at an MOI of 0.1. (B) The titre of virus in cells (keratinocytes, HSC-2, HSC-3, SCC-4, SAT, OK-92, Ca9-22, SKN-3, KON, KOSC-2, Sa-3, H-1, HO-1-N1, and HSC-4) was measured 96 h after reovirus infection at an MOI of 0.1.
Mentions: To examine whether the extent of cell cytotoxicity after SIN infection correlates with viral growth, viral titres in the cancer cell lines were measured 96 h after SIN or reovirus infection (0.1 MOI). High growth of SIN and reovirus was observed in all OSCC cell lines (Figure 2) whereas no viral growth was observed in normal keratinocytes, supporting the idea that the cytotoxicity caused by SIN depends on replication of the virus. However, there was little correlation between the viral titre and cytotoxicity caused by infection. For example, the SIN viral growth was high in HSC-4 cells but SIN infection caused little cytotoxicity in these cells. In contrast, viral growth was low in Sa-3 and H-1 cells but SIN caused high cytotoxicity in these cell lines.

Bottom Line: Sindbis viral growth was not observed in NHOKs, whereas high SIN growth was observed in all OSCC cell lines, including HCS-4.The CPE was shown, by terminal deoxyribonucleotidyl transferase-mediated dUTP nick-end labelling assays, to be apoptotic cell death.Furthermore, quantitative RT-PCR of mRNA in HSC-3 and HSC-4 cells after SIN infection showed that activation of caspases, cytochrome c, and IkappaBalpha was associated with SIN-induced apoptosis.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular Virology, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba 260-8670, Japan.

ABSTRACT

Background: Sindbis virus (SIN) infection causes no or only mild symptoms (fever, rash, and arthralgia) in humans. However, SIN has a strong cytopathic effect (CPE) on various cancer cells. This study focuses on the oncolytic activity of SIN AR399 on oral cancer cells compared with reovirus, a well-known oncolytic virus that targets cancer cells.

Methods: We analysed the cytotoxicity and growth of SIN in 13 oral squamous cell carcinoma (OSCC) cell lines (HSC-2, HSC-3, HSC-4, Ca9-22, H-1, Sa-3, KON, KOSC-2, OK-92, HO-1-N1, SCC-4, SAT, SKN-3) and normal human oral keratinocytes (NHOKs).

Results: Sindbis virus infection induced CPE in 12 OSCC cell lines at a low multiplicity of infection (MOI) of 0.01, but not in the OSCC cell line, HSC-4 or NHOKs. Sindbis viral growth was not observed in NHOKs, whereas high SIN growth was observed in all OSCC cell lines, including HCS-4. The cytotoxicity and growth of SIN was the same as reovirus at an MOI of 20 in 12 OSCC cell lines. The CPE was shown, by terminal deoxyribonucleotidyl transferase-mediated dUTP nick-end labelling assays, to be apoptotic cell death. Furthermore, quantitative RT-PCR of mRNA in HSC-3 and HSC-4 cells after SIN infection showed that activation of caspases, cytochrome c, and IkappaBalpha was associated with SIN-induced apoptosis.

Conclusion: As a replication-competent oncolytic virus, SIN may be a useful therapeutic modality for oral cancers.

Show MeSH
Related in: MedlinePlus