[Effects of survivin antisense RNA and HSP70 double gene transfection on MCF-7 cells in vitro].
OBJECTIVE
To explore the effects of survivin antisense RNA and HSP70 double gene transfection on breast cancer cell line MCF-7.


METHODS
MCF-7 cells was transfected with the double-gene vector pIRES2-EGFP-survivin antisense RNA/HSP70 via liposome. After a 72-h transfection, the cells were collected for observation under inverted fluorescent microscope. The changes of survivin mRNA and HSP70 protein expressions in the cells were detected with real-time PCR and Western-blot before and after the cell transfection, and the apoptotic rate of the transfected MCF-7 cells was detected by flow cytometry analysis with Annexin-V-cy5/7AAD double staining.


RESULTS
Green fluorescence was detected in MCF-7 cells transfected with the double-gene expression vector and the empty vector under inverted fluorescent microscope. The expression level of survivin mRNA in the cells was reduced effectively after the transfection with the double-gene expression vector, which also induced obvious cell apoptosis and enhanced the expression level of HSP70 protein as compared with those in MCF-7 cells transfected with the empty vector and the untransfected MCF-7 cells.


CONCLUSIONS
Survivin antisense RNA can interfere with the expression of endogenous survivin and induce apoptosis of MCF-7 cells. HSP70 can increase the expression of HSP70 protein in MCF-7 cells.
Gong
Dao-yuan
DY
Medical College, Foshan University of Science and Technology, Foshan 528000, China.
Guo
Ting-ting
TT
Fu
Hong-xia
HX
Peng
Yan
Y
Wang
Xiao-juan
XJ
D017209:Apoptosis / Q000187:drug effects*; D005260:Female; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics / Q000494:pharmacology*; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins / Q000235:genetics / Q000494:pharmacology*; D061986:MCF-7 Cells; D016372:RNA, Antisense / Q000235:genetics / Q000494:pharmacology*; D012333:RNA, Messenger / Q000235:genetics; D000077022:Survivin; D014162:Transfection
C107489:BIRC5 protein, human; D018840:HSP70 Heat-Shock Proteins; D051034:Inhibitor of Apoptosis Proteins; D016372:RNA, Antisense; D012333:RNA, Messenger; D000077022:Survivin