摘要
目的:利用miRNA21抑制物抑制食管癌细胞系ECA-109中miRNA21的表达,观察对ECA-109细胞凋亡的影响,为探讨miRNA21抑制物在食管癌治疗中的应用提供理论参考。方法:以正常培养的ECA-109细胞为空白对照组,转染miRNA21抑制物阴性对照物作为阴性对照组,转染miRNA21抑制物作为实验组,通过MTT法、流式细胞术、实时荧光定量PCR研究miRNA21抑制物对ECA-109细胞增殖、凋亡及凋亡相关基因表达的影响。结果:与空白对照组和阴性对照组相比,转染miRNA21抑制物能够显著性地抑制ECA-109细胞的增殖(P<0.05),并且明显提高ECA-109细胞的凋亡率(P<0.05),促进细胞凋亡。同时,转染miRNA21抑制剂的实验组ECA-109细胞中促凋亡基因caspase3、p53、在m RNA水平的表达均显著升高(P<0.05),而抗凋亡基因Bcl-2的表达则被显著抑制(P<0.05)。结论:miRNA21抑制剂能够抑制食管癌细胞ECA-109的增殖,增强促凋亡相关基因的表达,促进癌细胞的凋亡;miRNA21具有明显的原癌基因样功能,可作为食管癌细胞治疗的靶标。
Objective： The purpose of this study was to provide theoretical reference for the application of miRNA21 inhibitor in the treatment of esophageal carcinoma,by using miRNA21 inhibitor miRNA21 expression in ECA-109 esophageal cancer cell lines,to observe the influence of the ECA-109 cell apoptosis. Methods： The cultured ECA-109 cells were divided into blank control group（ normally cultured）,negative control group transfected with negative miRNA21 inhibitor,and the experimental group transfected with miRNA21 inhibitor. The influence of miRNA21 inhibitor on cell proliferation,apoptosis and apoptosis-related gene expression of ECA-109 cells was repectively investigated by MTT method,flow cytometry and real-time fluorescence quantitative PCR. Results： Compared with blank and negative control groups,the transfection of miRNA21 inhibitor significantly inhibited the proliferation of ECA-109 cells（ P〈0. 05）,significantly elevated the percent of apoptotic cells（ P〈0. 05） and promoted its apoptosis. While,the expression of pro-apoptotic genes as caspase3 and p53 was significantly elevated in ECA-109 cells of experimental group（ P〈0. 05）. However,anti-apoptotic gene Bcl-2 was significantly inhibited（ P〈0. 05）. Conclusion： miRNA21 inhibitors can inhibit the proliferation of esophageal cancer cell ECA-109,enhance the expression of pro-apoptotic genes and promote the apoptosis of esophageal carcinoma cells. Therefore,miRNA21 has the similar function of oncogene,which can be used as a target for the treatment of esophageal carcinoma cells.
出处
《川北医学院学报》
CAS
2017年第1期78-81,共4页
Journal of North Sichuan Medical College
