[Antisense oligonucleotide targeting survivin gene induces cell apoptosis in salivary mucoepidermoid carcinoma].
OBJECTIVE
to determine the effects of survivin antisense oligonucleotide (ASODN) on the expression levels of survivin mRNA and human mucoepidermoid carcinoma cell line (highly metastatic Mc3) apoptosis and to explore the feasibility of survivin gene as the mucoepidermoid carcinoma therapeutic targets.


METHODS
the survivin ASODN was designed and synthesized and then respectively transfected into Mc3 cells. The morphological changes of the Mc3 cells were observed 24, 48 and 72 h after transfection by inverted microscope and the apoptosis rate detected by flow cytometry. Methyl thiazolyl tetrazolium (MTT) assay was used to detect the effect of the transfection on cell poliferation, terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) method for analysis of apoptotic index, and semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) to detect the expression of survivin.


RESULTS
in survivin ASODN transfection group, there was less Mc3 cells than in other groups. The suspended cells dropping from the wall increased and showed typical apoptotic changes and time-dependent. Mc3 cell apoptosis rate in survivin ASODN transfection group transfected for 24, 48, 72 h was 12.96%, 14.43%, 22.69%, respectively, which were significantly higher than in other groups (P < 0.01) and time-dependent (P < 0.05). The inhibitory rate of Mc3 cells in survivin ASODN transfection group was 22.35%, 39.04%, 43.46%, which were significantly higher than other groups (P < 0.01) and time-dependent (P < 0.05). The apoptosis index (AI) of Mc3 cells in survivin ASODN transfection group was 11.038%, 12.172%, 18.900%, significantly higher than other groups (P < 0.01) and time-dependent (P < 0.05). The survivin mRNA levels in Mc3 cells were 0.739 ± 0.008, 0.668 ± 0.007, 0.500 ± 0.006, and the relative inhibition rate in these cells was 18.21%, 26.06%, 44.82%, significantly lower than other groups (P < 0.01) and time-dependent manner (P < 0.01).


CONCLUSIONS
survivin ASODN could inhibit the proliferation of Mc3 cells and induce the apoptosis of Mc3 cells. It also can inhibit the expression of survivin mRNA. Survivin can be used as a gene therapy targets for mucoepidermoid carcinoma.
Department of Oral Pathology, Xi'an Jiaotong University Stomatological Hospital, Xi'an 710004, China. qihong@mail.xjtu.edu.cn
Hong
H
Qi
Jian
J
Guo
Yin-Cheng
YC
Zahng
Shu-Wei
SW
Li
Li-Lin
LL
Yang
D017209:Apoptosis; D018277:Carcinoma, Mucoepidermoid / Q000378:metabolism*; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D018390:Gene Targeting; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins / Q000235:genetics*; D008869:Microtubule-Associated Proteins; D009363:Neoplasm Proteins; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; D012468:Salivary Gland Neoplasms / Q000378:metabolism*; D000077022:Survivin; D014162:Transfection
C107489:BIRC5 protein, human; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins; D009363:Neoplasm Proteins; D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; D000077022:Survivin