Gene amplification is clearly an important aspect of tumour growth and development and has prognostic significance in certain tumours. The identification and genetic characterisation of new areas of amplification in human malignancy remains an important goal in understanding the underlying genetic l
Identification of allelic loss on chromosome arm 6p in human astrocytomas by arbitrarily primed polymerase chain reaction
β Scribed by Yoshiki Saitoh; Janet M. Bruner; Victor A. Levin; Athanassios P. Kyritsis
- Publisher
- John Wiley and Sons
- Year
- 1998
- Tongue
- English
- Weight
- 152 KB
- Volume
- 22
- Category
- Article
- ISSN
- 1045-2257
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β¦ Synopsis
We employed the arbitrarily primed polymerase chain reaction (AP-PCR), which is a PCR-based genomic fingerprinting technique, to detect novel genetic alterations in human astrocytomas. DNA fingerprints generated by arbitrary primers were compared in normal lymphocytes and tumor tissues from the same individuals. We cloned and sequenced an AP-PCR band showing a greatly decreased intensity in tumor tissue DNA, relative to normal. Southern blot showed that this sequence was homozygously deleted in the tumor cell genome. Semiquantitative PCR analysis further showed significant decreases of signals in seven of 24 tumors, consistent with homozygous deletion of this sequence. The deleted sequence was localized to chromosome region 6p21.1 by the fluorescence in situ hybridization method. Microsatellite analysis also showed frequent allelic loss of this locus compared to neighboring loci in astrocytoma tissues. These results suggest the presence of a novel tumor suppressor gene at the 6p21.1 locus.
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DNA from 10 human glioma cell lines was analyzed by arbitrarily primed polymerase chain reaction. By fingerprinting of the DNA fragments obtained, the presence of fragment Qx with an abnormal signal was detected in one of the glioblastoma cell lines, CCF-STTG1. The nucleotide sequence of this fragme