鸟氨酸脱羧酶基因沉默对子宫内膜癌细胞Ishikawa的影响

目的利用RNA干扰技术,观察鸟氨酸脱羧酶（ornithine decarboxylase,ODC）基因在子宫内膜癌Ishikawa细胞的沉默效应及对Ishikawa细胞增殖周期的影响。方法设计合成以鸟氨酸脱羧酶基因为靶向目标的siRNA序列质粒,利用脂质体介导的方法将质粒转染至子宫内膜癌Ishikawa细胞,实验分三组：Ishikawa组,pSUPER-EGFP组,pSUPER-EGFP-ODC组。用Real-time PCR和Western blot检测ODC的表达情况。采用MTT和流式细胞术来检测ODC基因沉默对Ishikawa细胞生长的影响。结果 Real-time PCR和Western显示pSUPER-EGFP-ODC组细胞ODC mRNA和蛋白质表达水平明显下降。MTT示pSUPER-EGFP-ODC能显著抑制Ishikawa细胞的增殖活性,与Ishikawa相比,差异有高度统计学意义（P〈0.01）。流式细胞术结果示pSUPER-EGFP-ODC能显著阻滞Ishikawa细胞于G0/G1期,S期细胞数相应减少,并诱导细胞凋亡,与Ishikawa相比,差异有高度统计学意义（P〈0.01）。结论转染靶向ODC基因的siRNA序列质粒能够下调ODC基因的表达,抑制人子宫内膜癌细胞Ishikawa在体外的增殖,阻滞Ishikawa细胞于G0/G1期并诱导细胞凋亡。

Effect of Ornithine decarboxylase gene silencing on endometrial cancer Ishikawa

Objective To observe the effect of the ornithine decarboxylase gene silence on endometrial cancer Ishikawa cells by use the RNA interference technology.Methods The ornithine decarboxylase gene siRNA plasmid was designed and synthesized,and transfected into endometrial cancer Ishikawa cells,The experiment was divided into three groups: Ishikawa group,pSUPER-EGFP group,pSUPER-EGFP-ODC group.Real-time PCR and Western blot was used to detect the expression of ODC.MTT and flow cytometry were used to detect the impact of the ODC gene silencing on the growth of Ishikawa cells.Results Real-time PCR and Western results showed that the ODC mRNA and protein expression levels in pSUPER-EGFP-ODC group were significantly decreased.MTT results showed that the pSUPER-EGFP-ODC could significantly inhibit the proliferation activity of Ishikawa cells compared with the Ishikawa(P < 0.01).Flow cytometry results showed pSUPER-EGFP-ODC could significantly blocked of Ishikawa cells in G0/G1 phase,a corresponding reduction in the number of S-phase cells,and induction of apoptosis,compared with the Ishikawa(P < 0.01).Conclusion Transfected with targeting siRNA sequences of the ODC gene plasmid could down-regulate ODC gene expression,inhibit the proliferation of human endometrial carcinoma Ishikawa in vitro block of Ishikawa cells in G0/G1 phase and induction of apoptosis.