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目的:探讨Re1A反义寡核苷酸对喉癌Hep-2细胞增殖的影响。方法:设计Re1A反义寡核苷酸序列,转染Hep-2细胞后在不同的时间点行MTT检测瘤细胞生长抑制情况,取转染48h的Hep-2细胞行克隆形成实验,应用RT-PCR和Western blot检测RelA mRNA和蛋白的表达。结果:MTT显示转染Re1A反义寡核苷酸可明显抑制Hep-2细胞的增殖,随时间延长抑制率升高,与对照组比较差异有统计学意义。克隆形成实验显示Re1A反义寡核苷酸的Hep-2细胞克隆形成率明显低于对照组,与对照组比较差异有统计学意义。RT-PCR和Western blot结果显示Re1A反义寡核苷酸分别在mRNA和蛋白水平显著抑制RelA基因表达。结论:Re1A反义寡核苷酸可抑制Re1A的mRNA和蛋白的表达,并抑制Hep-2细胞增殖和克隆形成。
Objective: To investigate the effect of Re1A antisense oligonucleotide on the proliferation of laryngeal carcinoma Hep-2 cells. Methods: Re1A antisense oligonucleotide sequence was designed and transfected into Hep-2 cells. MTT assay was used to detect the growth inhibition of Hep-2 cells at different time points. Hep-2 cells transfected with Hep-2 cells for 48 h were assayed by RT- RelA mRNA and protein expression were detected by PCR and Western blot. Results: MTT showed that transfection of Re1A antisense oligonucleotide significantly inhibited the proliferation of Hep-2 cells. The inhibition rate of Hep-2 cells was increased with the increase of time. The difference was statistically significant compared with the control group. Clone formation experiments showed that the clonogenic rate of Hep-2 cells of Re1A antisense oligonucleotide was significantly lower than that of the control group, and the difference was statistically significant compared with the control group. The results of RT-PCR and Western blot showed that Re1A antisense oligonucleotide significantly inhibited RelA gene expression at mRNA and protein levels respectively. CONCLUSION: Re1A antisense oligonucleotide inhibits Re1A mRNA and protein expression and inhibits Hep-2 cell proliferation and colony formation.