甲状腺癌相关基因1在喉癌组织中的表达及对喉癌细胞增殖、迁移和侵袭的影响

The expression of thyroid cancer gene-1 in laryngeal cancer and its effect on proliferation,migration,and invasion of laryngeal cancer cells

目的探讨甲状腺癌相关基因1(thyroid cancer gene-1,TC-1)在喉癌组织中的表达及对喉癌细胞增殖、迁移和侵袭能力的影响。方法采用免疫组织化学方法检测48例喉癌患者的喉癌组织及对应癌旁正常喉黏膜中TC-1的表达水平。Lenti-GFP-Puro(对照质粒)、Lenti-TC-1-GFP-Puro、siRNA N-CTL(阴性对照)及siRNA TC-1转染喉癌细胞,分别命名为Control组、TC-1组、siRNA-Control组及siRNA-TC-1组。采用CCK-8法、划痕实验及Transwell实验检测喉癌细胞的增殖、迁移及侵袭能力。采用Western blot检测β-catenin、c-Myc及MMP-7蛋白表达的变化。结果TC-1在喉癌组织中的阳性表达率高于正常喉黏膜组织(χ^(2)=15.875,P<0.001)。喉癌组织中TC-1的表达强度与肿瘤部位(χ^(2)=8.261,P=0.041)、TNM临床分期(χ^(2)=8.308,P=0.040)及病理分级(χ^(2)=15.808,P=0.015)相关。与Control组比较,TC-1组喉癌细胞的增殖、迁移及侵袭能力增强(t=-4.671、-8.179、-26.130,P<0.001、P=0.004、P<0.001),c-Myc与MMP-7蛋白表达升高(t=-7.391、-4.793,P=0.018、0.041)。与siRNA-Control组比较,siRNA-TC-1组喉癌细胞的增殖、迁移及侵袭能力减弱(t=9.144、4.213、6.684,P<0.001、P=0.015、P=0.004),c-Myc与MMP-7蛋白表达降低(t=15.989、13.021,P=0.004、0.006)。结论TC-1在喉癌组织中呈高表达,其表达强度与肿瘤发生部位、TNM临床分期及病理分级相关。过表达TC-1或干扰TC-1的表达能增强或减弱喉癌细胞的增殖、迁移及侵袭能力,其机制可能为TC-1调节Wnt/β-catenin信号通路靶基因的表达有关。

OBJECTIVE To investigate the expression of thyroid cancer gene-1(TC-1)in laryngeal cancer and its influence on the proliferation,migration and invasion capacity of laryngeal cancer cells.METHODS The immunohistochemistry was used to detect the expression level of TC-1 in laryngeal carcinoma tissues and corresponding adjacent normal laryngeal mucosa of 48 patients with laryngeal carcinoma.The laryngeal carcinoma cells were transfected with Lenti-GFP-Puro(control plasmid),Lenti-TC-1-GFP-Puro,siRNA N-CTL(negative control)and siRNA TC-1,named group of Control,TC-1,siRNA-Control and siRNA-TC-1,respectively.The activity of proliferation,migration and invasion of laryngeal cancer cells were measured by CCK-8,scratch-wound and Transwell chamber assays,respectively.The expression level of protein aboutβ-catenin,c-Myc and MMP-7 were determined by Western blotting.RESULTS The positive expression rate of TC-1 in laryngeal carcinoma tissue was higher than that in normal laryngeal mucosa tissue(χ~2=15.875,P<0.001).The expression intensity of TC-1 in laryngeal cancer tissue was correlated with the tumor site(χ~2=8.261,P=0.041),advanced TNM stage(χ~2=8.308,P=0.040)and pathological grade(χ~2=15.808,P=0.015).Compared with the control group,there were increased in the proliferation,migration and invasion of laryngeal carcinoma cells(t=-4.671,-8.179,-26.130;P<0.001,=0.004,<0.001),and the protein expression levels of c-Myc and MMP-7 in the TC-1 group(t=-7.391,-4.793;P=0.018,0.041).Compared with the siRNA-Control group,there were decreased in the proliferation,migration and invasion of laryngeal carcinoma cells(t=9.144,4.213,6.684;P<0.001,P=0.015,P=0.004),and the protein expression levels of c-Myc and MMP-7 in the siRNA-TC-1group(t=15.989,13.021;P=0.004,0.006).CONCLUSION The expression of TC-1 was up-regulated in laryngeal cancer tissues,and its expression intensity was correlated with the tumor site,TNM stage and pathological grade.Overexpression of TC-1 or interference the expression of TC-1 can enhance or inhibit the proliferation,migration and invasion of laryngeal carcinoma cells.The mechanism may be that the TC-1 can regulate the expression of target genes of Wnt/β-catenin signaling pathway.