Higuchi Y, Kita K, Nakanishi H, Wang X L, Sugaya S, Tanzawa H, Yamamori H, Sugita K, Yamaura A, Suzuki N
Department of Neurosurgery, School of Medicine, Chiba University, Japan.
Biochem Biophys Res Commun. 1998 Jul 30;248(3):597-602. doi: 10.1006/bbrc.1998.8978.
UVr-10 UV-resistant cells were established from UV-sensitive human RSb cells. We searched here for genes expressed differentially between UVr-10 and RSb cells using a differential display method to elucidate the molecular mechanisms underlying the UV-resistance. Thirteen candidate cDNA fragments were obtained from 514 mRNA species first screened. Among the cDNA fragments, 3 revealed increased mRNA expression in UVr-10 cells compared with RSb cells by Northern analysis. Nucleotide sequence analysis identified 2 of the 3 cDNA fragments as encoding nucleophosmin (NPM) and T-plastin. The expression of NPM mRNA was induced after UV irradiation in UVr-10 cells but not in RSb cells, whereas irradiation did not affect the expression of T-plastin mRNA. UVr-10 cells transfected with antisense cDNA for NPM mRNA were partially sensitized to UV cell-killing. Thus, NPM and possibly T-plastin genes may contribute to the increased resistance to UV cell-killing, at least in the human cells tested.
UVr-10抗紫外线细胞系由对紫外线敏感的人RSb细胞构建而成。我们在此利用差异显示法寻找UVr-10细胞和RSb细胞之间差异表达的基因,以阐明抗紫外线能力的分子机制。从最初筛选的514种mRNA中获得了13个候选cDNA片段。通过Northern分析,在这些cDNA片段中,有3个显示UVr-10细胞中mRNA表达相对于RSb细胞增加。核苷酸序列分析确定这3个cDNA片段中的2个分别编码核仁磷酸蛋白(NPM)和T-肌动蛋白结合蛋白。UV照射后,UVr-10细胞中NPM mRNA的表达被诱导,而RSb细胞中则未被诱导,而照射对T-肌动蛋白结合蛋白mRNA的表达没有影响。用NPM mRNA反义cDNA转染的UVr-10细胞对紫外线诱导的细胞杀伤作用部分敏感。因此,至少在所测试的人类细胞中,NPM基因以及可能的T-肌动蛋白结合蛋白基因可能有助于增强对紫外线诱导的细胞杀伤作用的抗性。