成束蛋白、细胞角蛋白14在食管鳞癌中的表达

Expression of fascin and cytokeratin 14 in esophageal squamous cell carcinoma

目的检测和分析食管鳞癌组织、癌旁正常鳞状上皮及食管鳞癌细胞系中成束蛋白(fascin)和细胞角蛋白14(CK14)的表达情况,探讨这两种细胞骨架蛋白及相关蛋白在食管鳞癌发生发展中可能的作用及在食管鳞癌诊断中的应用前景。方法收集116例食管鳞癌组织标本,构建组织芯片。用免疫组化方法检测鳞癌组织与癌旁正常鳞状上皮中成束蛋白、CK14的表达情况,并分析它们与临床病理特征、预后的关系及两种蛋白之间的相关关系。用 Western 印迹方法检测12种食管鳞癌细胞系中成束蛋白、CK14的表达情况,并分析它们与生长和侵袭特点的关系。结果成束蛋白和CK14在食管正常鳞状上皮阴性(基底细胞除外),在食管鳞癌组织明显上调,阳性率分别为79.3%和67.0%,两者联合阳性率高达86.2%。成束蛋白和 CK14在高、中分化鳞癌中表达更高(成束蛋白在高、中、低分化鳞癌阳性率为87.1%、83.9%、62.1%,P=0.054;CK14在高、中、低分化鳞癌阳性率为87.1%、76.4%、27.6%,P<0.01)。两种蛋白表达与预后无明显关系(P=0.8980和 P=0.2610)。两种蛋白表达呈正相关(r=0.487,P<0.01)。成束蛋白在绝大多数细胞系高表达,仅在生长和侵袭能力弱的 TE12细胞系中无表达,而 CK14仅在 KYSE180细胞系中表达。结论成束蛋白、CK14在食管鳞癌普遍表达上调,可能在食管鳞癌发生发展中起重要作用。联合应用还有望成为食管鳞癌诊断的有效标志物。

Objective To investigate the expression of fascin and cytokeratin 14 （CK14） in human esophageal squamous cell carcinoma （ESCC） and the association of these two proteins with ESCC malignant progression and the possibility of application of these 2 proteins in the diagnosis of ESCC. Methods A tissue microarray composed of the representative regions of ESCC and corresponding normal epithelium was constructed. Immunohistochemistry was conducted to examine the expression of fascin and CD14 in 116 specimens of ESCC and the normal tissues near the cancerous tissues. The relation of these two proteins with the invasive depth, node involvement, differentiated grade, pTNM stages was analyzed. Disease-free survival analysis was carried out using Kaplan-Meier method. The correlation of the two proteins was analyzed using Spearman＇s correlation test. ESCC cells of the lines EC9706, TEl2, COLO-680N, KYSF.510, KYSE450, KYSE410, KYSE180, KYSE150, KYSE140, KYSE70, KYSE30, and YES2 were cultured and underwent SDS-PAGE and Western blotting to examine the expression of fascin and CD14. And the correlation of the two proteins with the characteristics of the cell lines was analyzed too. Results Fascin and CK14 were negative in the normal esophageal epithelia except in the basal cells. The positive rates of fascin and CK14 in the ESCC cells were 79. 3% and 67. 0% respectively. The positive rate of either fascin or CK14 was 86. 2%. The expression rates of fascin and CK14 in well- and moderately-differentiated ESCCs were significantly higher than that in the poorly-differentiated ones （P = 0. 054 and P 〈 0. 01 ）. The patients with positive expression of fascin and those with negative expression of CK14 had a poorer survival in comparison with those with negative fascin expression and those with positive CK14 expression respectively, however, without statistical significances （P = 0. 8980 and P = 0. 2610）. The positive rates of fascin in the well - , moderately- , and poorly - differentiated ESCCs were 87. 1% , 83.9% , and 62. 1% respectively （P = 0. 054）. The positive rates of CK14 in the well - , moderately - , and poorly - differentiated ESCCs were; 87. 1%, 76.4%, and 27.6% respectively （P 〈 0. 01 ）. The prognosis was not significantly correlated with the expression of both proteins （P = 0. 8980 and P = 0. 2610）. There was an significantly positive correlation between the expression levels of these 2 proteins （ r = 0. 487,P 〈 0. 01 ）. Fascin was highly expressed in most of the ESCC lines, except in the slowly growing and weakly migrating TE12 line. High expression of CK14 was only seen in the line KYSE180. Conclusion Fascin and CK14 may play important roles in the carcinogenesis and progression of ESCC. Combination of fascin and CK14 would be valuable markers in diagnosis of ESCC.