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[目的]研究RNA干扰抑制Skp2表达对喉癌鳞状细胞p27表达、细胞增殖和凋亡的作用。[方法]利用脂质体将重组质粒转染Hep-2细胞,用慢病毒系统建立干扰Skp2基团的稳定细胞株;荧光实时定量PCR和Western blot法检测转染细胞株中Skp2和p27m RNA及其蛋白表达;采用MTT法和流式细胞仪检测转染细胞的增殖和凋亡情况。[结果]通过慢病毒siRNA技术,Skp2可持续稳定抑制Hep2喉癌细胞,siRNA可诱导抑制Skp2,增加p27表达,降低细胞增殖,增加喉癌细胞的凋亡。[结论]靶向Skp2基因的si RNA对Hep-2细胞的抑制作用可能是通过上调p27基因水平来实现的,Skp2是基因疗法治疗喉癌的有利靶点。
[Objective] To investigate the effect of RNA interference on the expression of p27, proliferation and apoptosis of squamous cell carcinoma in laryngeal squamous cell carcinoma. [Method] The recombinant plasmids were transfected into Hep-2 cells by lipofectamine and the stable cell lines which interfered with Skp2 were constructed by lentivirus system. The expression of Skp2 and p27 mRNA in transfected cell lines was detected by real-time quantitative PCR and Western blot The protein expression of the transfected cells was detected by MTT assay and flow cytometry. [Result] Skp2 could inhibit Hep2 laryngeal carcinoma cells stably and stably by lentiviral siRNA. SiRNA could induce the inhibition of Skp2, increase the expression of p27, decrease the cell proliferation and increase the apoptosis of laryngeal carcinoma cells. [Conclusion] The inhibitory effect of si RNA targeting Skp2 gene on Hep-2 cells may be achieved by up-regulating the level of p27 gene. Skp2 is a good target for gene therapy for laryngeal cancer.