摘要
目的 :为了阐明人肺巨细胞癌高转移株 (95D)细胞高表达uPA、低转移株 (95C)细胞低表达uPA的原因 ,分析两株细胞中有关的转录因子的差异。方法 :uPA基因启动子不同片段与Luciferase报告基因的融合基因真核表达质粒分别平行转染 95C、95D细胞 ,比较活性 ;用EMSA分析 95D、95C细胞中与寡核苷酸探针特异结合的转录因子差异 ;用RT PCR分析两株细胞中相应转录因子mRNA相对表达水平。结果 :三个不同长度的uPA基因启动子在 95D细胞中的活性均相应高于 95C细胞中的活性 ,分别是 95C细胞的 1 32、1 6 6和 1 6 1倍 ,EMSA表明95D细胞中与AP1、AP2、NFκB及SP1探针结合的转录因子量均高于 95C细胞中 ;RT PCR表明AP2、NKκB及SP1转录因子mRNA表达水平分别是 95C细胞的 1 5 7、1 77和 1 2 8倍。结论 :转录因子AP1、AP2、NKκB及SP1在95D细胞中含量高于 95C细胞可能是uPA在 95D与 95C细胞中表达差别的原因之一。
Purpose:To study the difference of the transcription factors between the high(95D) and low(95C) metastatic human lung cancer cells which have different uPA expressions Methods:uPA gene promoter fragments of different length recombined with Luciferase reporter gene were transfected into 95C or 95D cells to analyse the different promoter activities between the two cells EMSA was performed to compare the transcription factors binding certain oligo nucleotides probes between the two cells RT PCR was used to realize the corresponding transcription factors mRNA relative expressions Results:Three different uPA gene promoter fragments activities are higher in 95D cell than that in 95C cell, being respectively 1 32, 1 66 and 1 61 times EMSA showed that transcription factors AP1, AP2, NKκB and SP1 contents are higher in 95D cell RT PCR suggested AP2, NKκB and SP1 transcription factors mRNA expressions are 1 57,1 77 and 1 28 times those in 95C cell Conclusions:The different uPA expressions between 95D and 95C cells are related to the higher transcription factors contents in 95D cell than in 95C cell
出处
《中国癌症杂志》
CAS
CSCD
2003年第5期418-422,共5页
China Oncology
基金
国家自然科学基金 (3 0 170 3 98)
关键词
尿激酶型纤溶酶原激活剂
人肺巨细胞癌
转录因子
高转移株
低转移株
表达
urokinase type plasminogen activator(uPA)
high and low metastatic human lung cancer cell lines
transcription factors
