Haugen A, Mann D, Murray C, Weston A, Willey J C
Laboratory of Human Carcinogenesis, National Cancer Institute, Bethesda, Maryland.
Mol Carcinog. 1989;2(2):68-71. doi: 10.1002/mc.2940020204.
Interleukin-1 alpha (IL-1 alpha) is a cytokine produced by a number of cell types including macrophages, fibroblasts, keratinocytes, and mesangial cells. We were interested in identifying a DNA restriction fragment length polymorphism (RFLP) for the IL-1 alpha gene for use in studies of genetic alteration in various human cancers. Human genomic DNA from 32 unrelated individuals was digested with various restriction enzymes, alone and in combination, and subjected to Southern blot analysis. Hybridization to 32P-labeled IL-1 alpha cDNA revealed an insertion-deletion-type polymorphic pattern. After digestion with RsaI, insertion-deletion-type polymorphic bands with sizes of 3.4 kb, 3.1 kb, and 2.8 kb and one invariant band of 0.8 kb were observed. These three alleles, designated A1, A2, and A3, had relative frequencies of 0.18, 0.06, and 0.78 with heterozygosity observed in 38% of the unrelated individuals studied. Evaluation of nine related individuals for this RsaI polymorphism was consistent with a Mendelian inheritance. Comparison of restriction patterns following Southern analysis of DNA digested with several different enzymes showed that the polymorphic region resides within the sixth intron. Furthermore, this RFLP results from a variable length region containing multiple copies of a recognition sequence for SP1, an imperfect copy of viral enhancer elements, and an inverse and complementary sequence of the glucocorticoid receptor binding site. The identified polymorphism may be of value in analyses of chromosome 2 and may help to elucidate mechanisms by which IL-1 alpha transcription is regulated.
白细胞介素-1α（IL-1α）是一种由多种细胞类型产生的细胞因子，包括巨噬细胞、成纤维细胞、角质形成细胞和系膜细胞。我们感兴趣的是鉴定IL-1α基因的DNA限制性片段长度多态性（RFLP），用于各种人类癌症的基因改变研究。从32名无亲缘关系的个体中提取的人类基因组DNA，用各种限制性酶单独或联合消化，然后进行Southern印迹分析。与32P标记的IL-1α cDNA杂交显示出插入-缺失型多态性模式。用RsaI消化后，观察到大小为3.4 kb、3.1 kb和2.8 kb的插入-缺失型多态性条带以及一条0.8 kb的不变条带。这三个等位基因，分别命名为A1、A2和A3，相对频率分别为0.18、0.06和0.78，在所研究的38%的无亲缘关系个体中观察到杂合性。对九名相关个体进行的这种RsaI多态性评估与孟德尔遗传一致。用几种不同酶消化DNA后进行Southern分析，比较限制性图谱表明多态性区域位于第六内含子内。此外，这种RFLP是由一个可变长度区域产生的，该区域包含SP1识别序列的多个拷贝、病毒增强子元件的不完全拷贝以及糖皮质激素受体结合位点的反向互补序列。所鉴定的多态性可能对2号染色体的分析有价值，并可能有助于阐明IL-1α转录调控的机制。
