TWIST介导的上皮-间质转化与宫颈癌发生发展相关性的研究

目的:探究TWIST介导的上皮间质化(EMT)在宫颈癌发生发展中的作用.方法:选取正常宫颈上皮、宫颈上皮内瘤变以及宫颈鳞状细胞癌患者标本共128例,石蜡包埋连续切片并运用免疫组织化学的方法检测TWIST和E-cadherin在各组织中表达情况；构建TWIST真核表达载体pcDNA3.1(+)/TWIST并建立高表达TWIST的宫颈癌细胞系pcDNA3.1(+)/TWIST/SiHa,观察稳转前后细胞形态学改变并运用蛋白质印迹法检测EMT相关标志蛋白变化.结果:TWIST在正常宫颈上皮、CIN Ⅰ～Ⅱ、CINⅢ到早期宫颈鳞癌(ⅠA～ⅡA期)及晚期宫颈癌(ⅡB～ⅢB期)中胞质阳性表达率分别为14.2％、63.4％、64.3％、100.0％和100.0％；胞核阳性表达率分别为21.4％、56.6％、57.1％、88.0％和100.0％.与此同时,E-cadherin在各期组织中胞膜表达阳性率分别为71.4％、36.7％、35.7％、12.0％和11.1％,而胞质表达阳性率分别为18.6％、63.3％、64.3％、88.0％和88.9％.同时,TWIST在各期组织胞质、胞核表达强度改变与E-cadherin在胞膜、胞质表达分布及强度改变相反.在伴有淋巴结转移的宫颈癌病例中,TWIST在胞质胞核中的强阳性率高于不伴淋巴结转移病例(P=0.012,P=0.033),高表达TWIST的SiHa细胞系细胞形态及EMT相关标志蛋白E-cadherin和Vimentin均发生明显改变,差异有统计学意义,P＜0.05.结论:TWIST在宫颈癌变发生发展的过程中表达逐渐升高,其介导的上皮间质转化贯穿宫颈癌发生发展的整个过程.

Correlation of epithelial-mesenchymal transitions induced by TWIST with tumorigenesis and progression of cervical cancer

OBJECTIVE:To investigate the relationship of TWIST induced epithelial-mesenchymal transitions and cervical cancer progression.METHODS:Immunohistochemistry was used to examine the distribution and expression of TWIST and E-cadherin in 128specimens inclnding normal cervial tissue,cervical intraepithelial neoplasia(CIN) and squamous cell carcinoma of cervix(SCC).To construct eukaryotic expression vector of human TWIST pcDNA3.1(+)/ TWIST and establish its stable transfected SiHa cell line.The morphology of the cell line was observed and alteration of the EMT markers was tested by western blot.RESULTS:In specimens of normal cervial tissues,CINⅠ-Ⅱ,CINⅢ,SCC(ⅠA-ⅡA),and SCC(ⅡB-ⅢB),the positive expression of TWIST in cytoplasm were 14.2%,63.4%,64.3%,100.0%,100.0%,and in cell nucleus were 21.4%,56.6%,57.1%,88.0%,100.0%.In the same specimens,the positive expression of E-cadherin in cellular membrane were 71.4%,36.7%,35.7%,12.0%,11.1%,and in cytoplasm were 18.6%,63.3%,64.3%,88.0%,88.9%.The inverse correlation was obtained between the expression of TWIST and E-cadherin.The expression of TWIST in the SCC groups with lymph node metastasis were higher than that in the groups without lymph node metastasis(P=0.012,P=0.033).The morphology of pcDNA3.1(+)/ TWIST / SiHa cell line obviously transformed and expression of EMT markers had also changed(P<0.05).CONCLUSIONS:The expression of TWIST escalats during the tumorigenesis and progression of cervical cancer.Additionally,EMT induced by TWIST plays an important role in this process.