[Antisense oligonucleotide targeting survivin induces apoptosis of renal clear-cell carcinoma cells and enhances their sensitivity to epirubicin in vitro].
OBJECTIVE
To investigate the effect of antisense oligonucleotide (ASODN) targeting survivin on the apoptosis and proliferation of renal cancer cell line 786-O and enhancement of its sensitivity to epirubicin.


METHODS
ASODN targeting survivin was designed and constructed. Cultured cells were divided into 6 groups: control group, liposome group, sense oligonucleotide (SODN) group, 600 nmol/L ASODN group, and 600 nmol/L ASODN combined with epirubicin group. After transfected for 24 h, cultured cells were harvested to carry on the next tests. Cell morphological changes were examined by transmission electron microscopy. Survivin protein was detected by immunohistochemical method. Apoptosis index (AI) and proliferation index (PI) were examined by flow cytometry.


RESULTS
Morphological abnormalities of cells were observed in ASODN transfected groups. Expression of survivin in ASODN groups were significantly decreased compared with that in the control group, liposomes group and SODN group. AI of ASODN groups was significantly higher than that in other groups. PI of ASODN groups was significantly lower than that in other groups. The PI of ASODN combined with epirubicin group was (35.7 +/- 1.67)%, but (9.3 +/- 0.34)% or (8.5 +/- 0.21)% in liposomes group or SODN group that had combined with epirubicin. The ASODN group achieved the strongest effects to enhance apoptosis in comparison with control group (P < 0.05), while SODN did not cause statistically significant change (P > 0.05).


CONCLUSIONS
The expression of survivin protein in the renal clear cell carcinoma cell line 786-O is downregulated by survivin ASODN. ASODN targeting survivin induces apoptosis and inhibits proliferation of 786-O cells. Inhibition of survivin enhances sensitivity of 786-O to epirubicin.
Gao
Xiao-kang
XK
Department of Urologic Surgery, Xijing Hospital, The Fourth Military Medical University, Xi'an 710032, China. gaoxkmn@fmmu.edu.cn
Li
Qing
Q
Wang
He
H
Yang
Bo
B
Yuan
Jian-lin
JL
Shao
Guo-xing
GX
Kang
Fu-xia
FX
D000903:Antibiotics, Antineoplastic / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D002292:Carcinoma, Renal Cell / Q000378:metabolism / Q000473:pathology*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D015251:Epirubicin / Q000494:pharmacology*; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins; D007680:Kidney Neoplasms / Q000378:metabolism / Q000473:pathology*; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000494:pharmacology*; D009363:Neoplasm Proteins / Q000235:genetics / Q000494:pharmacology*; D016376:Oligonucleotides, Antisense / Q000235:genetics / Q000494:pharmacology*; D000077022:Survivin; D014162:Transfection
D000903:Antibiotics, Antineoplastic; C107489:BIRC5 protein, human; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins; D009363:Neoplasm Proteins; D016376:Oligonucleotides, Antisense; D000077022:Survivin; D015251:Epirubicin