α-γ-干扰素诱导基因的差异表达

目的为分离转移相关基因,对两种干扰素诱导出现的差异表达ｃＤＮＡ片段进行序列测定。方法以α－和γ－干扰素分别处理ＭＡ８９１细胞,提取其ＲＮＡ,用Ｎｏｒｔｈｅｒｎｂｌｏｔ进一步确定用差异显示法所获得的基因片段的差异性。选择表达差异明显的片段进行ＤＮＡ测序。结果Ｎｏｒｔｈｅｒｎｂｌｏｔ结果表明,差异显示法得到的１０个ｃＤＮＡ片段中,４个确属差异表达片段。测序结果显示,Ｇ１５γ属尚无同源性报道的新基因;Ｃ４α与小鼠一种次要组织相容性复合体基因高度同源;Ｔ２γ与小鼠β－ａｃｔｉｎ基因高度同源;Ｔ１１γ与褐鼠高血压相关基因高度同源。结论确定了α－、γ－干扰素处理ＭＡ８９１细胞出现４条ｃＤＮＡ差异显示片段的核苷酸序列,为研究其与肿瘤转移的联系打下了基础。

IFN alpha- and IFN gamma-induced differentially expressed genes

OBJECTIVES: It was previously found that IFN alpha could reduce, while IFN gamma could enhance metastatic potential of a mouse mammary adenocarcinoma cell line MA891. The aim of this study was to identify the sequence of cDNA fragments that were differentially expressed by the two types of IFN, respectively. METHODS: Total RNA was extracted from MA891 cells treated with IFN alpha or IFN gamma. Northern blotting was used to confirm the differentially expressed gene fragments. DNA sequencing was then carried out. RESULTS: The Northern blots indicated that among the 10 cDNA fragments obtained by mRNA differential display technique, 4(G15 gamma, C4 alpha, T2 gamma, and T11 gamma) were shown to be unequivocally differentially expressed. Among them, the nucleotide sequence of G15 gamma did not show homology with any of the existing genes, while that of C4 alpha, T2 gamma and T11 gamma was highly homolgous to mouse minor histocompatility complex gene, mouse beta-actin gene and hamster high blood pressure gene, respectively. CONCLUSIONS: Four differentially expressed cDNA fragments by IFN alpha- and IFN gamma-treated MA891 cells have been ascertained. Regardless of sequence homology, none of them is reportedly related to tumor cell metastasis. The results herein reported provide basis for the elucidation of new metastasis-associated gene(s).