Monitoring Humans for Somatic Mutation in the Endogenous PIG-A Gene Using Red Blood Cells
Monitoring Humans for Somatic Mutation in the Endogenous PIG-A Gene Using Red Blood Cells
复制标题
DOI：
10.1002/em.20667
复制
发表时间：
2011-12-01
影响因子：
2.8
通讯作者：
Heflich, Robert H.
中科院分区：
文献类型：
--
作者：
Dobrovolsky, Vasily N.;Elespuru, Rosalie K.;Heflich, Robert H.
内源性X-连锁PIG-A基因参与糖基磷脂酰肌醇（GPI）锚的合成，所述锚将特异性蛋白质标记物拴系到哺乳动物细胞质膜的外部。在啮齿动物模型中的早期研究表明，在用遗传毒性剂处理的动物中可以诱导Pig-a突变红细胞（RBC），并且流式细胞术可以用于鉴定缺乏GPI锚定蛋白CD 59的罕见RBC，作为Pig-a基因突变的标志物。我们研究了类似的方法是否可以用于检测人类的基因突变。我们首先在97名自我鉴定的健康志愿者中确定了自发性CD 59缺陷型RBC（假定为PIG-A突变体）的频率。对于大多数受试者，CD 59缺陷型RBC的频率较低（平均值为5.1 +/- 4.9 x 10（-6）;中位数为3.8 x 10（-6），75%的受试者的突变频率低于8 x 10（-6）），男性和女性之间的中位突变频率存在统计学显著差异。PIG-A红细胞突变频率与年龄相关性较差，与吸烟状况无关。此外，两个个体的CD 59缺陷RBC频率显著增加，接近300 x 10（-6）和100 x 10（-6）。然后，我们监测了10名新诊断的癌症患者接受化疗与已知的遗传毒性药物的PIG-A突变。在化疗开始前测量患者血液中CD 59缺陷型RBC的频率，并在化疗期间/化疗后的6个月内测量3次。反应通常较弱，大多数观察结果均低于男性和女性的中位突变频率;最大反应是1例接受顺铂和依托泊苷联合治疗的患者中CD 59缺陷型RBC的频率增加约3倍。这些结果表明，RBC PIG-A试验可用于检测人体细胞突变。有必要进一步研究以确定该检测方法在检测通过不同机制起作用的遗传毒性剂诱导的突变方面的灵敏度。Environ.摩尔变异体52：784-794，2011.出版社：Wiley Periodicals，Inc.
The endogenous X-linked PIG-A gene is involved in the synthesis of glycosyl phosphatidyl inositol (GPI) anchors that tether specific protein markers to the exterior of mammalian cell cytoplasmic membranes. Earlier studies in rodent models indicate that Pig-a mutant red blood cells (RBCs) can be induced in animals treated with genotoxic agents, and that flow cytometry can be used to identify rare RBCs deficient in the GPI-anchored protein, CD59, as a marker of Pig-a gene mutation. We investigated if a similar approach could be used for detecting gene mutation in humans. We first determined the frequency of spontaneous CD59-deficient RBCs (presumed PIG-A mutants) in 97 self-identified healthy volunteers. For most subjects, the frequency of CD59-deficient RBCs was low (average of 5.1 +/- 4.9 x 10(-6); median of 3.8 x 10(-6) and mutant frequency less than 8 x 10(-6) for 75% of subjects), with a statistically significant difference in median mutant frequencies between males and females. PIG-A RBC mutant frequency displayed poor correlation with the age and no correlation with the smoking status of the subjects. Also, two individuals had markedly increased CD59-deficient RBC frequencies of similar to 300 x 10(-6) and similar to 100 x 10(-6). We then monitored PIG-A mutation in 10 newly diagnosed cancer patients undergoing chemotherapy with known genotoxic drugs. The frequency of CD59-deficient RBCs in the blood of the patients was measured before the start of chemotherapy and three times over a period of similar to 6 months while on/after chemotherapy. Responses were generally weak, most observations being less than the median mutant frequency for both males and females; the greatest response was an approximate three-fold increase in the frequency of CD59-deficient RBCs in one patient treated with a combination of cisplatin and etoposide. These results suggest that the RBC PIG-A assay can be adopted to measuring somatic cell mutation in humans. Further research is necessary to determine the assay's sensitivity in detecting mutations induced by genotoxic agents acting via different mechanisms. Environ. Mol. Mutagen. 52:784-794, 2011. Published 2011 Wiley Periodicals, Inc.