【摘要】 背景与目的:Fascin1蛋白为一种与F-肌动蛋白结合的55kDa细胞骨架蛋白,从人畸胎瘤中克隆fascin1基因。食管上皮细胞癌变机制至今不甚明了,为此,对永生化食管上皮细胞株SHEE(Shantouhumanembryonicesophagealepithelialcellline)和由SHEE恶性转化而来的食管癌细胞株SHEEmt之间的差异表达基因在蛋白质和mRNA两个水平上进行检测分析,以期为揭示食管鳞状上皮细胞癌变机制提供新思路。方法:采用双向凝胶电泳技术分析SHEE与SHEEmt之间的蛋白质差异表达情况,以基质辅助激光解吸附电离飞行时间质谱(matrix-assistedlaserdesorption/ionizationtimeofflyingmassspectrometry,MALDI-TOF-MS)对部分差异表达蛋白质点进行鉴定,以逆转录聚合酶链反应(reversetranscriptionploymerasechainreaction,RT-PCR)对目标基因进行mRNA转录水平上的检测,并对RT-PCR产物进行序列测定分析。结果:在SHEE向SHEEmt的恶性转化中有9个基因出现差异表达,其中fascin1基因在蛋白质水平升高3.64倍,mRNA水平升高16.17倍。结论:fascin1基因可能与SHEE向SHEEmt的恶性转化有关。更多还原

【Abstract】 BACKGROUND & OBJECTIVE: Fascin 1 is the 55kDa F-actin- binding cytoskeleton protein. Fascin 1 gene was cloned from a human teratocarcinoma. Up to now, the carcinogenesis mechanism of esophageal squamous cell carcinoma is un clear. The study was designed to identify the differentially expressed proteins and mRNAs between the human immortalized esophageal epithelial cell line (SHEE) transfected by human papillomavirus type18 E6E7 and the malignant transformati on cell line (SHEEmt), which is derivated from SHEE, and to further understand t he carcinogenesis mechanisms of esophageal squamous cell carcinoma. METHODS: Cel lular proteins were separated by two-dimensional electrophoresis and differenti ally expressed proteins were identified by matrix-assisted laser desorption/ion ization time of flying mass spectrometry (MALDI-TOF-MS). The mRNA of fascin 1 gene was assayed by reverse transcription ploymerase chain reaction (RT-PCR) an d its product was analyzed by sequencing assay. RESULTS: The results manifested 9 genes expressed differently in the progress of malignant transformation of SH EE to SHEEmt,fascin 1 protein increased about 3.64 times and its mRNA increased about 16.17 times. CONCLUSION: The upregulated expression of fascin 1 gene may b e correlated with the malignant transformation of SHEE to SHEEmt.更多还原