[Induction of apoptosis and inhibition of proliferation in Hep-2 by survivin gene targeting in vitro].
OBJECTIVE
To study induction of apoptosis and inhibition of proliferation in Hep-2 by survivin gene targeting.


METHODS
Antisense survivin RNA expression vector was constructed and then was transfected to human laryngeal carcinoma cell line Hep-2 by lipofectamine. HpEGFP/survivin cells were abstained by using G418. The level of survivin protein before and after transfection was determined by Western blot. Proliferation activity was measured by MTT assay. The experiment of colony formation in soft agar was carried out. Apoptosis was assessed by flow cytometry and acridine orange (AO).


RESULTS
After antisense survivin RNA plasmids were transfected, the level of survivin protein was inhibited in Hep-2. Compared with control, proliferation of HpEGFP/survivin cells transfected with the recombinant of antisense survivin RNA were suppressed significantly. The experiment of colony formation in soft agar showed the ability of colony formation decreased in HpEGFP/survivin cells compared to control (P <0.05). Apoptosis rate increased about 1.81 folds compared with control.


CONCLUSIONS
The recombination antisense survivin RNA plasmid can partly inhibit the level of survivin protein expression in Hep-2. Antisense survivin RNA can induce apoptosis and inhibit the proliferation of Hep-2 by down-regulating the expression of endogenous survivin in vitro.
Wen
Lianji
L
Department of Otorhinolaryngology, the Second Hospital of Jilin University, Changchun, China.
Li
Changqing
C
Li
Ye
Y
Zhao
Dan
D
Gao
Lifang
L
Wang
Lihua
L
Guan
Guofang
G
Jin
Chunshun
C
D017209:Apoptosis; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D018390:Gene Targeting; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins; D007822:Laryngeal Neoplasms / Q000473:pathology; D008869:Microtubule-Associated Proteins / Q000235:genetics*; D010957:Plasmids; D016372:RNA, Antisense / Q000235:genetics; D000077022:Survivin; D014162:Transfection
C107489:BIRC5 protein, human; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins; D016372:RNA, Antisense; D000077022:Survivin