张大伟, 吴剑, 任德莲, 等.CLDN1在食管鳞癌中的表达及其分布异常在食管癌发生中的作用.四川大学学报(医学版),2017,48(6):850-856
CLDN1在食管鳞癌中的表达及其分布异常在食管癌发生中的作用
Expression of CLDN1 and Its Nuclear Distribution in Esophageal Squamous Carcinoma
  
中文关键词:  CLDN1食管鳞癌肿瘤发生
英文关键词:CLDN1Esophageal squamous carcinomaTumorigenesis
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中文摘要:
      目的 检测紧密连接蛋白CLDN1在食管鳞癌组织中的表达,研究CLDN1对食管鳞癌TE-11细胞生物学功能的影响。方法 应用免疫组化检测食管鳞癌组织芯片(含30例食管鳞癌组织、30例癌旁组织及30例食管远端组织)中CLDN1的表达,比较食管癌组织及癌旁组织中CLDN1的表达差异;应用Western blot检测15例食管鳞癌患者术后组织和食管癌细胞株CLDN1的表达,分析CLDN1在肿瘤细胞及正常食管上皮细胞内的分布;构建包含CLDN1 shRNA序列的慢病毒并转染TE-11细胞,采用Western blot和流式细胞术检测CLDN1干扰效果及细胞内分布。通过CCK-8检测细胞增殖能力;Transwell检测细胞侵袭迁移能力;采用Rhodamine phalloidin染色TE-11细胞骨架并在激光共聚焦显微镜下观察细胞肌丝变化。结果 免疫组化结果显示CLDN1在癌组织及癌旁组织中阳性表达率明显差异( P>0.05),但在高分化和中分化的食管鳞癌组织CLDN1呈强阳性表达,低分化食管鳞癌组织CLDN1表达明显降低;Western blot检测CLDN1在TE-11细胞中主要分布在细胞核内;干扰CLDN1表达后,TE-11细胞增殖明显减慢,侵袭及迁移能力降低。激光共聚焦显微镜显示下调CLDN1表达后,TE-11细胞骨架蛋白荧光强度减弱,细胞表面肌丝减少。结论 CLDN1在食管鳞癌组织中的表达与其分化程度相关,在TE-11中具有促癌作用,其表达及分布异常与肿瘤的发生发展密切相关,有望作为食管鳞癌的重要生物标志物。
英文摘要:
      ObjectiveTo determine the expression of tight junction protein CLDN1 in esophageal squamous carcinoma (ESCC) and to explore the effect of CLDN1 on the biological function of ESCC TE-11 cells. MethodsThis study collected 30 ESCC tissues, 30 adjacent normal tissues and 30 distal esophageal tissues to detected the expression of CLDN1 in tissue microaary by immunohistochemistry method. Western blot was employed to determine the expression of CLDN1 in 15 ESCC tissues and cell lines, and then the distribution of CLDN1 in tumor cells and normal esophageal epithelial cells was analyzed. Lentivirus containing the sequence of CLDN1 shRNA was constructed and transfected into TE-11 cells, the transfection efficiency and intracellular distribution was determined by Western blot and flow cytometry. Cell proliferation was detected by CCK-8, the invasion and migration ability of cells was determined with Transwell. Cytoskeleton changes was observed by laser scanning confocal microscope. ResultsImmunohistochemical results showed no significant difference between the cancer tissues and adjacent tissues in term of the positive rate of CLDN1 ( P>0.05). CLDN1 was highly expressed in highly and moderately differentiated ESCC tissues, but its expression was significantly decreased in poorly differentiated cancer tissues. Western blot revealed that CLDN1 was mainly distributed in the nucleus of TE-11 cells. The proliferation of TE-11 cells, as well as invasion and migration ability, were declined obviously when the expression of CLDN1 being down-regulated, while the cytoskeleton protein fluorescence intensity deceased.ConclusionThe expression of CLDN1 in ESCC tissue is associated with its differentiation and may promote carcinogenesis in TE-11 cells.
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