[The effect of cell killing and apoptosis by human herpes simplex virus- thymidine kinase/ganciclovir system combined with allitride in BIU87 cells].
OBJECTIVE
To study the killing effect of human herpes simplex virus-thymidine kinase/ganciclovir (HSV-TK/GCV) system combined with allitride and the possible apoptosis mechanism in BIU87 cells.


METHODS
The cytotoxicity after combination were estimated by theamine blue tetrazolium bromide (MTT). The morphological changes were observed with inverted microscope and in-situ cell apoptosis detection kit. Changes of apoptosis rate and cell cycle were assessed by flow cytometry. B-cell lymphoma-2 (bcl-2), bax, caspase-3 (cysteine aspartate specific proteinase) mRNA changes were detected by reverse transcriptase polymerase chain reaction, and caspase-3 activity was estimated with colorimetry.


RESULTS
For combination group, the cell killing rate was raised to 72.50% to compare with 35.00% of GCV and 37.00% of allitride separately and there was a synergistic effect between these two drugs. The cell apoptosis was induced in all three groups and for the combination group the time of S-phase and G(2)-phase arrest were earlier than other two groups. Both drugs could inhibit the expression of bcl-2 and promote the expression and activity of caspase-3.


CONCLUSIONS
The combination of HSV-TK/GCV system with allitride can inhibit the proliferation of BIU87 cells congenerously through apoptosis, which may be correlated with S- and G(2)-phase arrest, down-regulation of bcl-2 and increased caspase-3 expression and its activity.
Qiu
Shao-peng
SP
Department of Urology, First Affiliated Hospital of Sun Yat-sen University, Guangzhou 510080, China. urology@vip.163.com
Mao
Xiao-peng
XP
Cao
Kai-yuan
KY
Chen
Xian-jing
XJ
Yuan
Guang-qing
GQ
Xu
Lin
L
Huang
Xiao-rong
XR
D017209:Apoptosis / Q000187:drug effects; D004220:Disulfides; D004357:Drug Synergism; D015774:Ganciclovir / Q000494:pharmacology*; D015316:Genetic Therapy; D018259:Herpesvirus 1, Human / Q000201:enzymology / Q000235:genetics*; D006801:Humans; D066298:In Vitro Techniques; D013441:Sulfinic Acids / Q000494:pharmacology*; D013937:Thymidine Kinase / Q000235:genetics*; D014162:Transfection; D001749:Urinary Bladder Neoplasms / Q000473:pathology* / Q000628:therapy
D004220:Disulfides; D013441:Sulfinic Acids; C006452:allicin; D013937:Thymidine Kinase; D015774:Ganciclovir