[Antisense RNA of survivin gene enhances the taxol-induced apoptosis and sensitivity to chemotherapy drugs in bladder cancer cells: an in vitro experiment].
OBJECTIVE
To evaluate the effects of antisense oligodeoxynucleotide (ASODN) of survivin gene on taxol-induced apoptosis in bladder cancer cells.


METHODS
A survivin ASODN eukaryotic vector pcDNA3-SVVas was transfected into human bladder cancer cells of the line BIU87 mediated by liposomal reagent (BIU87 SVVas cells). The mRNA expression of survivin was measured by RT-PCR. Blank plasmid pcDNA3 was transfected as control (BIU87 neo cells). The cell growth curve was drawn using trypan blue dye exclusion assay. MTT assay was used to investigate the sensibility of transfected cells to taxol. The BIU87 SVVas cells, BIU87 neo cells, and BIU87 cells were cultured in the culture fluid with taxol and then DNA gel electrophoresis, Hoechst nuclear staining and fluorescent microscopy, and annexin-propidium iodide (PI) staining were used to examine the cell apoptosis.


RESULTS
Compared to BIU87 and BIU87 neo cells, the mRNA expression of survivin in the BIU87-SVVas cells was obviously reduced. The growth of the BIU87-SVVas cells was significantly reduced in comparison with the BIU87 (P < 0.05). The sensibility of BIU87 SVVas cells to taxol increased significantly shown by cell proliferation and MTT assays. Agarose gel electrophoresis of genomic DNA showed typical DNA ladder only in the BIU87 SVVas cells, but not in other cells. Hoechst nuclear staining and fluorescent microscopy showed that the nuclei of the BIU87 SVVas cells become condense. Annexin-PI test showed that the cell apoptosis rate of the BIU87-SVVas cells treated with taxol was significantly higher than those of the other groups.


CONCLUSIONS
survivin antisense RNA enhances the taxol-induced apoptosis in the bladder cancer cells. This may lay an experimental foundation for further research of biotherapy in bladder cancer.
Wang
Zhi-Hua
ZH
Department of Urology, Tongji Hospital, Tongji Medical College of Huazhong University of Sciences and Technology, Wuhan 430030, China.
Ye
Qin
Q
Huang
De-Ying
DY
Hu
Zhi-Quan
ZQ
Ye
Zhang-Qun
ZQ
Yang
Nuo
N
Liu
Hao
H
Zhuang
Qian-Yuan
QY
Yang
Wei-Min
WM
Liu
Ji-Hong
JH
Zhang
Xu
X
D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects* / Q000235:genetics; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects / Q000235:genetics; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins / Q000235:genetics*; D009363:Neoplasm Proteins / Q000235:genetics*; D017239:Paclitaxel / Q000494:pharmacology*; D016372:RNA, Antisense / Q000235:genetics*; D000077022:Survivin; D014162:Transfection; D001749:Urinary Bladder Neoplasms / Q000188:drug therapy / Q000235:genetics / Q000473:pathology
D000972:Antineoplastic Agents, Phytogenic; C107489:BIRC5 protein, human; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins; D009363:Neoplasm Proteins; D016372:RNA, Antisense; D000077022:Survivin; D017239:Paclitaxel