【摘 要】
：
为了研究反义封闭NGAL基因表达对SHEEC食管癌细胞微丝骨架以及肿瘤细胞生物学行为的影响,以不同长度NGAL基因片段反义表达载体和硫代修饰反义寡核苷酸单链片段转染SHEEC食管
【机 构】
：
汕头大学医学院中心实验室,汕头大学医学院肿瘤病理研究室,汕头大学医学院生物化学与分子生物学教研室,中国预防医学科学院病毒学研究所
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为了研究反义封闭NGAL基因表达对SHEEC食管癌细胞微丝骨架以及肿瘤细胞生物学行为的影响,以不同长度NGAL基因片段反义表达载体和硫代修饰反义寡核苷酸单链片段转染SHEEC食管癌细胞,通过C418筛选,建立一系列旨在封闭SHEEC食管癌细胞NGAL基因表达的亚细胞克隆.在细胞内F-肌动蛋白(F-actin)及DNA荧光双标记基础上,通过流式细胞术、激光共聚焦显微镜扫描术等技术手段检测封闭反义NGAL基因表达后,SHEEC食管癌细胞中F-actin和DNA含量、F-actin形态结构以及肿瘤细胞生物学行为的变化特征.结果显示,反义封闭NGAL基因表达后,SHEEC食管癌细胞F-actin的含量明显降低,与永生化食管上皮细胞SHEE相近,但细胞分裂增殖指数未见明显变化.表明反义封闭NGAL基因表达对SHEEC食管癌细胞的微丝骨架有明显影响,而对SHEEC食管癌细胞的分裂增殖影响不明显.激光共聚焦显微镜扫描观测显示,反义封闭NGAL基因表达可使SHEEC食管癌细胞F-actin分布均匀,F-actin小体减少,细胞间连接重新建立,结构较紧密,主要形态结构特征与SHEE细胞趋于一致.提示反义封闭NGAL基因表达可对SHEEc食管癌细胞的微丝骨架F-actin产生明显影响,推测癌细胞的微丝骨架F-actin可能是NGAL基因在SHEEC食管癌细胞中发挥功能的一种作用环节.
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