短发夹RNA沉默长链非编码RNA HULC对喉癌细胞生长和运动能力以及裸鼠成瘤的影响

Effect of shRNA HULC on growth and motor ability of laryngeal carcinoma cells and tumor formation in nude mice

目的:探究sh-HULC对喉癌细胞生长和运动能力以及裸鼠成瘤的影响。方法:构建shRNA HULC载体转染至Hep-2细胞,将Hep-2细胞随机分为3组:Control组、shRNA-NC组、sh-HULC组。EDU染色检测细胞增殖;Transwell检测细胞侵袭;蛋白免疫印迹检测Ki67、Survivin、VEGF、Vimentin蛋白表达水平;构建裸鼠移植瘤模型,将裸鼠随机分为两组:Control组和shHULC组,检测肿瘤体积,实时荧光定量PCR(qPCR)检测HULC水平,免疫组化检测Ki67、VEGF阳性表达率。结果:与Control组比较,sh-HULC组EDU阳性细胞数,Ki67、Survivin蛋白水平、侵袭细胞数,VEGF、Vimentin蛋白水平均显著降低(P<0.05);与Control组比较,sh-HULC组肿瘤体积第21、第24、第27、第30天显著降低,HULC水平及Ki67、VEGF阳性表达率均显著降低(P<0.05)。结论:sh-HULC通过抑制细胞增殖、侵袭降低Hep-2细胞生长和运动能力,并抑制裸鼠肿瘤生长。

Objective: To investigate the effects of sh-HULC on the growth and motor ability of laryngeal carcinoma cells and tumor formation in nude mice. Methods: shRNA HULC vector was constructed and transfected into Hep-2 cells. Hep-2 cells were randomly divided into 3 groups: control group, shRNA-NC group and SHHULC group. Cell proliferation was detected by EDU staining. Transwell detected cell invasion. Protein expression levels of Ki67, Survivin, VEGF, and Vimentin were detected by Western blotting. Nude mice were randomly divided into two groups: control group and sh-HULC group. Tumor volume was detected. HULC level was detected by qPCR, and the positive expression rates of Ki67 and VEGF were detected by immunohistochemistry.Results: Compared with the control group, the number of EDU positive cells andinvaded cells, the levels of Ki67, Survivin VEGF and Vimentin proteins in the sh-HULC group were significantly decreased(P<0.05). In the nude mouse transplantation tumor experiment, compared with the control group, tumor volume of sh-HULC group was significantly reduced on day 21, 24, 27 and 30. HULC level was significantly reduced, and the positive expression rates of Ki67 and VEGF were significantly reduced(P<0.05). Conclusion: sh-HULC reduces the growth and motor ability of Hep-2 cells and inhibits tumor growth in nude mice by inhibiting cell proliferation and invasion.