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小鼠大细胞淋巴瘤中转移相关细胞表面糖蛋白和mRNA的差异表达

Differential expression of metastasis-associated cell surface glycoproteins and mRNA in a murine large cell lymphoma.

作者信息

Nicolson G L, LaBiche R A, Frazier M L, Blick M, Tressler R J, Reading C L, Irimura T, Rotter V

出版信息

J Cell Biochem. 1986;31(4):305-12. doi: 10.1002/jcb.240310408.

Abstract

A metastatic variant cell subline of the Abelson virus-transformed murine large lymphoma/lymphosarcoma RAW117 has been selected in vivo ten times for liver colonization. Highly metastatic subline RAW117-H10 forms greater than 200 times as many gross surface liver tumor nodules as the parental line RAW117-P. Analysis of cellular proteins and glycoproteins indicates reduced expression of murine Moloney leukemia virus-associated p15, p30, and gp70, and increased expression of a sialoglycoprotein, gp150, in the highly metastatic H10 cells. Northern analyses of oncogene expression suggested that mRNA of various oncogenes was expressed equally or not expressed in the RAW117 cells of differing metastatic potential. Differential gene expression was examined using a cDNA library of 17,600 clones established from poly A+ mRNA isolated from H10 cells. The cDNA library was screened by the colony hybridization technique using probes made from both RAW117-P and -H10 cells. Approximately 99.5% of these cDNA clones were expressed identically in P and H10 cells. Of the few differentially expressed cDNA clones (approx. 150/17,600), one-half of these were identified as Moloney leukemia virus sequences in a separate probing with a radiolabeled Moloney leukemia virus probe. The remainder of the differentially expressed mRNA detected by colony hybridization of the cDNA library were expressed at higher levels (approx. 1/6) or lower levels (approx. 1/3) in the highly metastatic H10 cells.

摘要

从经阿贝尔森病毒转化的小鼠大型淋巴瘤/淋巴肉瘤RAW117中筛选出一个转移变异细胞亚系,该亚系已在体内进行了十次肝脏定植筛选。高转移亚系RAW117-H10形成的肝脏表面肉眼可见肿瘤结节数量比亲代系RAW117-P多200倍以上。对细胞蛋白质和糖蛋白的分析表明,在高转移的H10细胞中,小鼠莫洛尼白血病病毒相关的p15、p30和gp70表达降低,而一种唾液酸糖蛋白gp150表达增加。对癌基因表达的Northern分析表明,各种癌基因的mRNA在具有不同转移潜能的RAW117细胞中表达水平相同或未表达。使用从H10细胞分离的聚腺苷酸加尾mRNA构建的包含17600个克隆的cDNA文库,对差异基因表达进行了检测。使用从RAW117-P和-H10细胞制备的探针,通过菌落杂交技术对该cDNA文库进行筛选。这些cDNA克隆中约99.5%在P细胞和H10细胞中的表达相同。在少数差异表达的cDNA克隆中(约150/17600),其中一半在使用放射性标记的莫洛尼白血病病毒探针单独探测时被鉴定为莫洛尼白血病病毒序列。通过cDNA文库菌落杂交检测到的其余差异表达mRNA在高转移的H10细胞中表达水平较高(约1/6)或较低(约1/3)。

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