Batra S K, Metzgar R S, Hollingsworth M A
Department of Microbiology and Immunology, Duke University Medical Center, Durham, North Carolina 27710.
J Biol Chem. 1991 Apr 15;266(11):6830-3.
A cDNA library from a poorly differentiated human pancreatic tumor cell line was screened for differentially expressed mRNAs using single-stranded cDNA probes synthesized from poly(A+) RNA of the poorly differentiated cell line Panc 1 and a very well differentiated cell line CD11. One of the cDNA clones isolated hybridized to a transcript size of 650 base pairs on Northern blot analysis and showed 30-fold higher expression in the poorly differentiated cell line as compared with the well differentiated cell line. Sequence analysis of this cDNA clone and its deduced amino acid sequence showed an open reading frame of 441 nucleotides with 100 and 98.6% homology to ribosomal protein S16 (rpS16) from rat and mouse, respectively. Northern blot analyses with a panel of 14 pancreatic cell lines, 2 breast cell lines, 2 colon cell lines, and several other tissues showed higher expression of rpS16 only in the poorly differentiated pancreatic tumor cell line Panc 1. The expression of mRNA for two other ribosomal proteins, rpL30 and rpL32, were not elevated in Panc 1. Southern blot analysis of genomic DNA showed a 20-fold amplification of a single band among the rpS16 family only in the Panc 1 cell line.
利用从低分化人胰腺肿瘤细胞系Panc 1和高分化细胞系CD11的聚腺苷酸(poly(A+))RNA合成的单链cDNA探针,对低分化人胰腺肿瘤细胞系的cDNA文库进行差异表达mRNA的筛选。分离得到的一个cDNA克隆在Northern印迹分析中与650个碱基对大小的转录本杂交,并且在低分化细胞系中的表达比高分化细胞系高30倍。对该cDNA克隆及其推导的氨基酸序列进行序列分析,显示一个441个核苷酸的开放阅读框,与大鼠和小鼠的核糖体蛋白S16(rpS16)分别具有100%和98.6%的同源性。用一组14个胰腺细胞系、2个乳腺细胞系、2个结肠细胞系和其他几种组织进行Northern印迹分析,结果显示仅在低分化胰腺肿瘤细胞系Panc 1中rpS16表达较高。另外两种核糖体蛋白rpL30和rpL32的mRNA表达在Panc 1中未升高。基因组DNA的Southern印迹分析显示,仅在Panc 1细胞系中rpS16家族中的一条带发生了20倍的扩增。