MUTATION OF P53 IN PRIMARY BIOPSY MATERIAL AND CELL-LINES FROM HODGKIN DISEASE

被引:50
作者
GUPTA, RK
PATEL, K
BODMER, WF
BODMER, JG
机构
[1] ST BARTHOLOMEWS HOSP,IMPERIAL CANC RES FUND,DEPT MED ONCOL,LONDON EC1A 7BE,ENGLAND
[2] IMPERIAL CANC RES FUND,HUMAN CYTOGENET LABS,LONDON WC2A 3PX,ENGLAND
[3] IMPERIAL CANC RES FUND,CANC GENET LAB,LONDON WC2A 3PX,ENGLAND
关键词
REED-STERNBERG CELLS; SINGLE-STRAND CONFORMATION POLYMORPHISM; SEQUENCING; IMMUNOCYTOCHEMISTRY; INSITU HYBRIDIZATION;
D O I
10.1073/pnas.90.7.2817
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
The p53 tumor-suppressor gene encodes a nuclear phosphoprotein that arrests cell cycle progress at G1. It may facilitate DNA damage repair and is frequently mutated in many human tumors. Hodgkin disease, a malignant condition of the lymphoid system, is characterized by the presence of Reed-Sternberg cells and mononuclear variants (Hodgkin cells), whose etiology remains unknown. The large multinucleated Reed-Sternberg cells often comprise <1% of the total cell population within a biopsy specimen and are thought to be the neoplastic component in an admixture of reactive cells. It has been shown in the large majority of cases that up to 60% of these multinucleated cells react with CM-1, an anti-p53 antibody. However, whether this ''overexpression'' of p53 protein reflects abnormality at the DNA level can no longer be assumed by immunocytochemistry alone. p53 from six Hodgkin disease-derived cell lines was examined by immunoprecipitation, polymerase chain reaction (PCR)-single-strand conformation polymorphism analysis, and sequencing. In one cell line, point mutations were identified in exons 5 and 8 of p53. Sequencing of cloned PCR products confirmed the mutations to be on different alleles. A strategy involving extraction of nuclei followed by enrichment by flow cytometry was used to determine whether p53 overexpression in the Reed-Sternberg cells from patient biopsy material was due to mutations in this gene. Single-strand conformation polymorphism revealed additional bands in the polyploid nuclear preparations, suggesting abnormalities, and sequence analysis confirmed the presence of point mutations.
引用
收藏
页码:2817 / 2821
页数:5
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