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[Antisense oligonucleotide targeting survivin gene induces cell apoptosis in salivary mucoepidermoid carcinoma]
Hong Qi1, Jian Guo, Yin-Cheng Zahng
1Department of Oral Pathology, Xi'an Jiaotong University Stomatological Hospital, Xi'an 710004, China. qihong@mail.xjtu.edu.cn
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.
Insights
Survivin antisense oligonucleotide (ASODN) effectively reduces survivin mRNA levels, inhibits cancer cell proliferation, and induces apoptosis in mucoepidermoid carcinoma cells. These findings support survivin as a viable gene therapy target for this cancer.
Area of Science:
- Oncology
- Molecular Biology
- Gene Therapy
Background:
- Mucoepidermoid carcinoma is a type of salivary gland cancer.
- Survivin is a protein that plays a role in inhibiting apoptosis and promoting cell proliferation.
- Targeting survivin presents a potential therapeutic strategy for various cancers.
Purpose of the Study:
- To investigate the effects of survivin antisense oligonucleotide (ASODN) on survivin mRNA expression and apoptosis in human mucoepidermoid carcinoma cells (Mc3).
- To evaluate the potential of the survivin gene as a therapeutic target for mucoepidermoid carcinoma.
Main Methods:
- Survivin ASODN was synthesized and transfected into Mc3 cells.
- Cell morphology, apoptosis rates (flow cytometry, TUNEL assay), proliferation (MTT assay), and survivin mRNA levels (RT-PCR) were assessed at various time points post-transfection.
Main Results:
- ASODN transfection led to significant increases in Mc3 cell apoptosis rates and apoptosis index in a time-dependent manner.
- Cell proliferation was significantly inhibited by ASODN transfection.
- Survivin mRNA expression levels were significantly reduced in ASODN-treated cells.
Conclusions:
- Survivin ASODN effectively inhibits proliferation and induces apoptosis in mucoepidermoid carcinoma cells.
- ASODN treatment significantly downregulates survivin mRNA expression.
- Survivin represents a promising gene therapeutic target for mucoepidermoid carcinoma.
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