[关键词]
[摘要]
目的：观察血小板源性生长因子受体α反义寡核苷酸(platelet-derived growth factor receptor-α antisense oligonucleotide，PDGFR-α ASODN)对体外培养兔晶状体上皮细胞N/N1003A增殖的影响，并探讨其作用机制。
方法：使用阳离子脂质体LipofectamineTM2000将PDGFR-α ASODN转染入N/N1003A细胞，MTT法分析细胞的增殖活性，RT-PCR法检测PDGFR-αmRNA 的表达，透射电镜观察细胞超微结构改变，流式细胞仪检测细胞的周期和凋亡率。
结果：PDGFR-α ASODN处理后，N/N1003A细胞增殖明显受到抑制(P<0.05)，PDGFR-α mRNA表达明显下调(P<0.05)，均呈剂量依赖性； 透射电镜观察到细胞出现典型的凋亡特征； 细胞凋亡率显著升高(P<0.05)，同时细胞阻滞于G1 期。
结论：PDGFR-α ASODN可沉默兔晶状体上皮细胞中PDGFR-α基因表达，抑制细胞增殖，并诱导细胞凋亡，为应用PDGFR-α ASODN预防后囊膜混浊的形成提供实验依据。
[Key word]
[Abstract]
AIM:To observe the effect of platelet-derived growth factor receptor-α antisense oligonucleotide(PDGFR-α ASODN)on the proliferation of rabbit lens epithelial cells N/N1003A
in vitro and investigate its mechanism.
METHODS: PDGFR-α ASODN was transfected into N/N1003A cells with LipofectamineTM 2000. The proliferations of the transfected cells were analyzed by MTT assay and the expressions of PDGFR-α mRNA were detected by RT-PCR. Phase microscope and transmission electron microscope were used to observe the changes of cell morphology and ultramicrostructure. Flow cytometry was applied to detect the changes of cell cycle and apoptosis rate.
RESULTS: After N/N1003A cells were treated by PDGFR-α ASODN, the proliferations of the cells were significantly inhibited(P<0.05), and the expression levels of PDGFR-α mRNA were notablely lowered(P<0.05)in a dose-dependent manner, respectively. The cells in the experimental group showed typical features of apoptosis under the transmission electron microscope and the rates of cell apoptosis were markedly higher(P<0.05). The analysis of cell cycle indicated that the cells were arrested in G1 phase.
CONCLUSION: PDGFR-α ASODN can silence PDGFR-α gene expression in rabbit lens epithelial cells, inhibit cell proliferation, and induce cell apoptosis, which provides the experimental basis for the application of PDGFR-α ASODN to prevent the formation of posterior capsular opacification.
[中图分类号]
[基金项目]
中国广西科学基金项目(No. 0899004)
