Expression of beta-actin during progression of mouse skin tumors.
Expression of beta-actin during progression of mouse skin tumors.
复制标题
小鼠皮肤肿瘤进展过程中β-肌动蛋白的表达。
DOI：
10.1093/carcin/10.8.1439
复制
发表时间：
1989
期刊：
影响因子：
4.7
通讯作者：
Bowden,GT
中科院分区：
文献类型：
--
作者：
Ostrowski,LE;Krieg,P;Finch,J;Cress,AE;Nagle,RB;Bowden,GT
重组DNA技术已被用来分离小鼠皮肤多阶段癌变过程中过度表达的序列。这六个 cDNA 序列之一 mal-4 检测到 1.9 kb 转录物，其在鳞状细胞癌 (SCC) 中的表达水平是正常表皮的 10 倍。 mal-4 的全长 cDNA 是从由产生 SCC 的细胞系 PDVC57 制成的 λgt10 cDNA 文库中获得的。对 mal-4 cDNA 的 1900 bp 中的 1300 bp 进行 DNA 测序并搜索 Genbank，结果显示整个 cDNA 与小鼠β-肌动蛋白的 DNA 序列相似性大于 95%，并且 3' 非翻译区的相似性大于 98%，这是各种肌动蛋白亚型所独有的。 Southern分析表明，在肿瘤进展过程中，β-肌动蛋白基因没有发生扩增或重排。 RNA:RNA 混合保护测定用于筛选小鼠皮肤肿瘤中突变β-肌动蛋白的表达。在任何检查的肿瘤中都没有获得突变的证据。用罗丹明结合的鬼笔环肽染色的肿瘤切片的荧光显微镜显示出 F-肌动蛋白定位的外周模式，乳头状瘤和癌之间没有明显差异。对从正常表皮、乳头状瘤或鳞状细胞中提取的蛋白质进行二维凝胶电泳后，观察到大约等量的β-肌动蛋白，表明鳞状细胞中β-肌动蛋白RNA的过度表达不会导致β-肌动蛋白稳态水平增加。这些结果表明肌动蛋白代谢的改变伴随着肿瘤的进展。
Recombinant DNA techniques have been employed to isolate sequences that are over-expressed during multi-stage carcinogenesis in the mouse skin. One of these six cDNA sequences, mal-4, detected 1.9-kb transcripts that were expressed at levels 10-fold higher in squamous cell carcinomas (SCCs) in comparison to normal epidermis. A full-length cDNA for mal-4 was obtained from a λgt10 cDNA library made from an SCC-producing cell line, PDVC57. DNA sequencing of 1300 out of 1900 bp of the mal-4 cDNA and searching of Genbank showed >95% DNA sequence similarity to mouseβ-actin over the entire cDNA and >98% similarity over the 3' untranslated region, which is unique to the various isoforms of actins. Southern analysis showed no amplification or rearrangement of theβ-actin gene had occurred during tumor progression. The RNA:RNA hybrid protection assay was utilized to screen for the expression of mutatedβ-actin(s) in mouse skin tumors. No evidence for mutation was obtained in any of the tumors examined. Fluorescence microscopy of tumor sections stained with rhodamine-conjugated phalloidin showed a peripheral pattern of F-actin localization with no gross differences between papillomas and carcinomas. Approximately equal amounts ofβ-actin were observed following two-dimensional gel electrophoresis of proteins extracted from normal epidermis, papillomas or SCCs, indicating that the over-expression ofβ-actin RNA in SCCs did not result in an increased steadystate level ofβ-actin protein. These results suggest that alterations in actin metabolism accompany tumor progression.