Okaichi K, Wang L H, Ihara M, Okumura Y
Department of Radiation Biophysics, Nagasaki University School of Medicine, Japan.
J Radiat Res. 1998 Jun;39(2):111-8. doi: 10.1269/jrr.39.111.
We have constructed an in vitro system to examine how p53 mutants affect radiosensitivity. Mutations of p53 were made using in vitro mutagenesis, and mutant cDNAs were introduced into the human osteosarcoma cell line, Saos-2, which is devoid of endogenous p53. For wild type p53, both the expression plasmid and a regulation plasmid (LacSwitch system) were transfected into the cells. The radiosensitivities of clones of mutant p53 and wild type p53 were examined. Transformants of wild type p53 had increased radiosensitivity. The induction of wild type p53 protein by addition of IPTG did not significantly increased radiosensitivity. A mutation at codon 123 also increased radiosensitivity. Mutations at codons 143, 175, and 273 did not alter radiosensitivity.
我们构建了一个体外系统来研究p53突变体如何影响放射敏感性。使用体外诱变技术制造p53突变,然后将突变的cDNA导入缺乏内源性p53的人骨肉瘤细胞系Saos-2中。对于野生型p53,将表达质粒和调控质粒(LacSwitch系统)都转染到细胞中。检测了突变型p53和野生型p53克隆的放射敏感性。野生型p53的转化体放射敏感性增加。添加IPTG诱导野生型p53蛋白并未显著增加放射敏感性。密码子123处的突变也增加了放射敏感性。密码子143、175和273处的突变未改变放射敏感性。