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KMID : 1001020110090020055
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Journal of Urologic Oncology
2011 Volume.9 No. 2 p.55 ~ p.61
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Anticancer Effects of Conditionally Replicating Oncolytic Herpes Simplex Virus Against Human Bladder Cancer Cell Lines
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Joo Kwan-Joong
Ko Kwang-Chon
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Abstract
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Purpose: We constructed oncolytic virus from herpes simplex virus(HSV), which selectively targets the activated Ras signaling pathway in cancer cells. We evaluated the anticancer effects of the oncolytic HSV against human bladder cancer cell lines.
Materials and Methods: Oncolytic HSV type 2(FusOn-H2) was constructed by replacing protein kinase domain of the ICP10 with green fluorescent protein gene. We seeded T24, 5637, TCC-SUP, and RT4 cell lines into 24 well plates respectively and infected them with FusOn-H2 at 0.1 and 0.01 plaque forming unit(pfu)/cell, or left them uninfected. Fluorescence microscopic examination was performed to characterize phenotypical finding after incubation for two days. Cells were harvested 24, 48 or 72 hours later and the number of viable cells determined after Trypan blue staining. The percentage of viable cells was calculated by dividing the number of cells excluding Trypan blue in the infected well by the number excluding the stain in the uninfected well.
Results: In FusOn-H2 infection, syncytia formation of dead cells was present and green fluorescence expression was observed. In all cell lines, infection with FusOn-H2 killed the tumor cells more and more as the time goes by. By 72 hours, FusOn-H2 almost completely eradicated the tumor cells exposed to virus doses of 0.1pfu/cell. There was no difference in cell destruction by oncolytic HSV between coxsackie-adenovirus receptor(CAR) expression cell lines(5637 and RT4) and CAR deficiency cell lines(T24 and TCC-SUP).
Conslusions: Oncolytic HSV has potent anticancer activity against bladder cancer irrespective of CAR expression. Probably, the treatment using oncolytic HSV in bladder cancer patients would lead to therapeutic benefits.
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KEYWORD
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Urinary bladder neoplasms, Cell line, Tumor, Oncolytic virotherapy, Simplexvirus
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