Xia F, Liber H L
Department of Cancer Biology, Harvard University School of Public Health, Boston, MA 02115, USA.
Mutat Res. 1997 Jan 3;373(1):87-97. doi: 10.1016/s0027-5107(96)00193-5.
Abnormalities in the p53 gene play an important role in genomic instability and tumorigenesis. Our previous work showed that p53 status is correlated with differential mutability in two closely related human lymphoblastoid cell lines, TK6 and WTK1. WTK1 cells, which contain a mutation in p53 (p53Ile237) show a remarkably increased mutability, larger genetic alterations at the thymidine kinase locus (tk), an increased ability to catalyze recombination, and a delay in the onset of apoptosis after X-irradiation, compared to TK6 (p53 +/+). In the present study, we demonstrate that after transfection and subsequent overexpression of the known dominant negative mutant p53 Ala143 allele (mp53Ala143) in TK6, there were significantly enhanced spontaneous and X-ray-induced mutant frequencies at the tk locus, and delayed onset of X-ray-induced apoptosis, to a similar extent as in WTK1. In addition, high protein expression of mp53Ala143 in transfectants was correlated with both increased mutation frequency and altered apoptosis kinetics. Similar results were obtained with p53 Ile237 transfection into TK6. Our observations indicate that the product of the p53 gene affects mutational processes. We hypothesize that p53 dysfunction can lead to increased mutagenicity at the endogenous tk gene in human lymphoblastoid cell lines either through delayed apoptosis in response to DNA damage or by mediating increased recombination.
p53基因的异常在基因组不稳定和肿瘤发生中起重要作用。我们之前的研究表明,p53状态与两种密切相关的人类淋巴母细胞系TK6和WTK1中的差异突变性相关。与TK6(p53 +/+)相比,含有p53突变(p53Ile237)的WTK1细胞显示出显著增加的突变性、胸苷激酶基因座(tk)处更大的基因改变、催化重组的能力增强以及X射线照射后凋亡起始延迟。在本研究中,我们证明在TK6中转染并随后过表达已知的显性负性突变体p53 Ala143等位基因(mp53Ala143)后,tk基因座处的自发和X射线诱导的突变频率显著增加,并且X射线诱导的凋亡起始延迟,程度与WTK1相似。此外,转染细胞中mp53Ala143的高蛋白表达与突变频率增加和凋亡动力学改变均相关。将p53 Ile237转染到TK6中也获得了类似的结果。我们的观察结果表明,p53基因的产物影响突变过程。我们假设p53功能障碍可通过对DNA损伤的凋亡延迟或通过介导重组增加导致人类淋巴母细胞系内源性tk基因的致突变性增加。