ZEB1的表达与肺鳞状细胞癌侵袭及转移的关系

Relationship between ZEB1 expression and invasion and metastasis of lung squamous cell carcinoma

目的:探讨锌指E-盒结合同源异形盒-1(ZEB1)在肺鳞状细胞癌组织中的表达,分析ZEB1mRNA和蛋白的表达水平与肺鳞状细胞癌侵袭及转移的关系。方法:选择45例肺鳞状细胞癌行外科手术切除的患者,根据所取组织与肿瘤的距离不同分为肺鳞状细胞癌组织和癌旁正常肺组织,采用RT-PCR和Western blotting方法检测肺鳞状细胞癌组织和癌旁正常肺组织中ZEB1mRNA和蛋白的表达情况,并分析肺鳞状细胞癌不同临床病理特征ZEB1mRNA和蛋白的表达差异;构建ZEB1特异性siRNA载体,感染肺癌NCI-H2170细胞,设未转染对照组、转染对照组和ZEB1siRNA转染组,应用Western blotting方法从蛋白质水平检测各组干扰质粒对ZEB1基因的干扰效果,通过Transwell侵袭小室观察各组NCI-H2170细胞侵袭能力的变化。结果:ZEB1mRNA在肺鳞状细胞癌组织中的阳性表达率显著高于癌旁正常肺组织(P<0.05);在淋巴结转移、远处转移阳性者肺癌组织中的阳性表达率显著高于其在淋巴结转移、远处转移阴性者肺癌组织中的表达(P<0.05);但ZEB1mRNA的表达与肺鳞状细胞癌患者的性别、年龄及肿瘤大小及分化程度无关(P>0.05)。肺鳞状细胞癌组织中ZEB1蛋白的表达与ZEB1mRNA的表达结果类似,ZEB1蛋白在肺鳞状细胞癌组织中的表达量显著高于癌旁正常肺组织(P<0.05);在淋巴结转移和远处转移阳性者癌组织中的表达显著高于其在淋巴结转移和远处转移阴性者癌组织中的表达(P<0.05)。下调NCI-H2170细胞中ZEB1蛋白的表达后,NCI-H2170细胞的侵袭及转移能力明显降低(P<0.05)。结论:ZEB1在肺鳞状细胞癌组织中高表达,其表达变化与肺鳞状细胞癌的病理特征密切相关;运用RNA干扰技术有效沉默NCI-H2170细胞的ZEB1基因后,可显著降低NCI-H2170细胞的侵袭及转移能力,提示ZEB1在肺癌的发生发展中起重要作用,抑制ZEB1的表达可能成为一种治疗肺癌的方法。

Objective To investigate the expression of zinc finger E-box binding homeobox 1（ZEB1） in lung squamous cell carcinoma,and to analyze the relationship between the ZEB1 mRNA and protein expression levels and invasion and metastasis of lung squamous cell carcinoma.Methods 45 patients with lung squamous cell carcinoma performed surgical operation excision were chosen.According to the distance from the tumor tissue,lung squamous cell carcinoma tissue and adjacent normal lung tissue were selected,RT-PCR and Western blotting were used to detect the ZEB1 mRNA and protein expressions in lung squamous cell carcinoma and adjacent normal lung tissues,and the relationship between the clinical pathological features of lung squamous cell carcinoma and ZEB1 mRNA and protein expressions were analyzed.ZEB1 specific siRNA vector was constructed,then NCI-H2170 cells were infected,meanwhile untransfected group,control group and ZEB1siRNA transfected group were set up.Western blotting was performed to detect the interference effect,and then through the Transwell chamber the changes of NCI-H2170 cell invasion ability were observed.Results The positive mRNA expression rate of ZEB1 in lung squamous cell carcinoma tissues was significantly higher than that in normal lung tissue（P〈0.05）.The positive mRNA expression rates in the carcinoma tissues from the patients with positive lymph node or distant metastasis were significantly higher than those from the patients without lymph node or distant metastasis（P〈0.05）.However,the expression of ZEB1 had no correlation with gender,age,tumor size,or tumor differentiation level of the patients with lung squamous cell carcinoma（P〉0.05）.The protein expression of ZEB1 was similar to the mRNA expression of ZEB1.The protein expression of ZEB1 in lung squamous cell carcinoma tissue was significantly higher than that in normal lung tissue（P〈0.05）.The protein expression level of ZEB1 in the carcinoma tissue from the patients with positive lymph nodes or distant metastasis was significantly higher than that from the patients without lymph nodes or distant metastasis（P〈0.05）.Reducing the protein expression of ZEB1 in NCI-H2170 cells,the number of cells that passed through the Transwell chamber was significantly lower than that in non-transfected control group as well as transfected control group（P〈0.05）.Conclusion ZEB1 displays a high expression in lung squamous cell carcinoma tissue,and their expression changes are associated with the pathological features of lung squamous cell carcinoma;By RNA interference effectively silencing ZEB1 gene in NCI-H2170 cells,knockdown of ZEB1 could significantly reduce the invasion and metastasis ability of NCI-H2170 cells,suggesting that ZEB1 play an important role in lung carcinogenesis and development,and inhibition of ZEB1 expression might become a kind of method for treating lung cancer.