Kohno T, Morishita K, Takano H, Shapiro D N, Yokota J
National Cancer Center Research Institute, Tokyo, Japan.
Oncogene. 1994 Jan;9(1):103-8.
We have searched for novel genetic alterations in human cancer cell lines by using the arbitrarily primed polymerase chain reaction (AP-PCR), which is a PCR-based genomic fingerprinting. A homozygous deletion was detected in a small-cell lung carcinoma (SCLC) cell line, NCI-H82. Since homozygous deletion is a critical genetic alteration for the inactivation of tumor suppressor gene, we defined the locus of homozygous deletion. Fluorescence in situ hybridization analysis revealed that the deletion was localized at chromosome 2q33. Allelic loss on chromosome 2q was detected in 29.4% (5/17) of SCLC and 37.5% (12/32) one non-SCLC by restriction fragment length polymorphism analysis. Considerably high incidence of allelic loss on chromosome 2q was also detected in colorectal carcinoma and neuroblastoma. These results suggest the presence of a novel tumor suppressor gene at 2q33, which is involved in the development of several human cancers. The size of the homozygous deletion in the NCI-H82 cell line was more than 20 kilobase pairs. Seven loci mapped to 2q32-qter were all retained in this cell line, suggesting the presence of submicroscopic interstitial deletion in this cell line. Homozygous deletion detected in this study should be an invaluable tool for positional cloning of the target tumor suppressor gene at 2q33.
我们通过使用任意引物聚合酶链反应（AP-PCR）来搜索人类癌细胞系中的新型基因改变，AP-PCR是一种基于PCR的基因组指纹技术。在小细胞肺癌（SCLC）细胞系NCI-H82中检测到纯合缺失。由于纯合缺失是肿瘤抑制基因失活的关键基因改变，我们确定了纯合缺失的位点。荧光原位杂交分析显示该缺失定位于染色体2q33。通过限制性片段长度多态性分析，在29.4%（5/17）的SCLC和37.5%（12/32）的非SCLC中检测到染色体2q上的等位基因缺失。在结直肠癌和神经母细胞瘤中也检测到染色体2q上等位基因缺失的发生率相当高。这些结果表明在2q33存在一个新的肿瘤抑制基因，它参与了几种人类癌症的发生发展。NCI-H82细胞系中纯合缺失的大小超过20千碱基对。定位于2q32-qter的7个位点在该细胞系中均得以保留，表明该细胞系中存在亚显微间隙缺失。本研究中检测到的纯合缺失对于在2q33位置克隆目标肿瘤抑制基因应是一种非常有价值的工具。
