siRNA干扰C-C趋化因子配体5对喉癌细胞生物学特性的影响

Effect of siRNA interference with C-C chemokine ligand 5 on biological characteristics of laryngeal carcinoma cells

目的研究C-C趋化因子配体5(C-C chemokine ligand 5,CCL5)在头颈鳞状细胞癌中的表达,并探讨CCL5对喉癌细胞生物学特性的影响。方法基因表达谱数据动态分析(Gene Expression Profiling Interactive Analysis,GEPIA)数据库查找CCL5在头颈鳞状细胞癌中的表达情况。将喉癌细胞TU177转染siRNA(siRNA组),并设对照(NC)组,CCK8法、细胞划痕试验、流式细胞术分别检测各组细胞增殖、迁移、细胞周期及细胞凋亡情况;RT-PCR、Western blot法检测CCL5敲降效率及多药耐药蛋白2(multidrug resistance protein 2,MRP2)、bcl-2相关x蛋白(bcl-2-associated x protein,Bax)mRNA转录及蛋白表达水平。结果CCL5在头颈鳞状细胞癌的表达高于正常组织(P<0.05)。与NC组相比,siRNA组siRNA具有较高的敲降效率(t分别为12.898和22.656,P均<0.01);siRNA敲降CCL5可抑制喉癌细胞增殖、迁移,促进喉癌细胞晚期凋亡及凋亡蛋白Bax的表达(t=2.600~11.667,P均<0.05)。结论CCL5在头颈鳞状细胞癌中高表达,siRNA干扰CCL5通过上调Bax表达抑制喉癌细胞TU177增殖、迁移,促进凋亡,为CCL5可能作为治疗喉癌的新靶点奠定了基础。

Objective To investigate the expression of C-C chemokine ligand 5(CCL5)in head and neck squamous cell carcinoma(HNSCC),and explore the effect of CCL5 on the biological characteristics of laryngeal carcinoma cells.Methods Gene Expression Profiling Interactive Analysis(GEPIA)database was used to investigate the expression of CCL5 in HNSCC.The laryngeal carcinoma cells TU177 were transfected with siRNA(siRNA group),and the control(NC)group was set up.The cell proliferation,migration,cycle and apoptosis of each group were detected by CCK8 assay,cell scratch test and flow cytometry respectively.RT-PCR and Western blot were used to detect the knock-down efficiency of CCL5 and the mRNA transcription and protein expression of multidrug resistance protein 2(MRP2)and bcl-2-associated x protein(Bax).Results The expression of CCL5 in HNSCC was higher than that in normal tissues(P<0.05).Compared with NC group,siRNA showed higher knock-down efficiency(t=12.898 and 22.656 respectively,each P<0.01);siRNA interference with CCL5 inhibited the proliferation and migration of laryngeal carcinoma cells,and promoted the late apoptosis of laryngeal carcinoma cells and the expression of apoptosis protein Bax(t=2.600~11.667,each P<0.05).Conclusion CCL5 was highly expressed in HNSCC,while siRNA interference with CCL5 inhibited the proliferation,migration and promoted apoptosis of laryngeal carcinoma cells TU177 by up-regulating the expression of Bax,which laid a foundation of the possibility of CCL5 as a new target for the treatment of laryngeal carcinoma.