Infection with oncolytic herpes simplex virus-1 induces apoptosis in neighboring human cancer cells: a potential target to increase anticancer activity.
OBJECTIVE
The antitumor efficacy of a herpes simplex virus (HSV)-1 oncolytic virus depends on the cytotoxic effect of the virus, but also on viral replication and spread within the tumor. Apoptosis is considered a defense mechanism of infected cells that minimizes the spread of viral progeny by limiting cellular production of virus. We sought to determine whether oncolytic HSV-1 infection induces apoptosis in neighboring, uninfected cells and whether manipulation of apoptosis can increase viral replication and cytotoxicity.


METHODS
NV1066 is an oncolytic HSV-1 mutant that contains the marker gene for enhanced green fluorescent protein. OCUM human gastric cancer cells were infected with NV1066 in vitro and inspected for apoptosis by Hoechst and terminal deoxynucleotidyltransferase-mediated nick end labeling staining and for infection by expression of green fluorescence.


RESULTS
A significant increase in apoptosis was seen in cells infected by NV1066. More interestingly, a significant percentage (10%) of uninfected cells also proceeded to apoptosis. After NV1066 infection, cells were also treated with N-acetylcysteine (NAC), an inhibitor of apoptosis. By day 4 after infection, 2.7x more NV1066 was produced in cells exposed to NAC than in those not exposed to NV1066 (P = 0.04). NAC also increased tumor kill when administered with virus.


CONCLUSIONS
These data suggest that NV1066 induces apoptosis in uninfected cocultured cells, potentially hindering propagation of viral progeny and concomitant tumor kill. Inhibition of apoptosis may improve the efficacy of oncolytic HSV-1 therapy.
Stanziale
Stephen F
SF
Department of Surgery, Hepatobiliary Division, Memorial Sloan-Kettering Cancer Center, New York, New York 10021, USA.
Petrowsky
Henrik
H
Adusumilli
Prasad S
PS
Ben-Porat
Leah
L
Gonen
Mithat
M
Fong
Yuman
Y
D000111:Acetylcysteine / Q000494:pharmacology; D017209:Apoptosis; D045744:Cell Line, Tumor; D005434:Flow Cytometry; D005770:Gastrointestinal Neoplasms / Q000473:pathology / Q000517:prevention & control / Q000821:virology; D015870:Gene Expression / Q000187:drug effects; D049452:Green Fluorescent Proteins; D018259:Herpesvirus 1, Human / Q000235:genetics / Q000254:growth & development*; D006801:Humans; D020287:In Situ Nick-End Labeling; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins; D000111:Acetylcysteine