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目的探讨60Coγ射线照射前、后,BRCA1、RAD51蛋白在GM、ATM+-AT、AT细胞中的表达。方法应用免疫荧光染色和激光扫描共聚焦显微镜,观察GM、ATM+-AT、AT细胞在0和10Gy60Coγ射线照射后,BRCA1、RAD51蛋白在上述细胞中的定位及表达并进行定量分析。结果60Coγ射线照射前,BRCA1、RAD51蛋白在GM、ATM+-AT、AT细胞中仅有少量表达并且无共定位表达,其中在AT细胞中表达量最低;10Gy60Coγ射线照射后,BRCA1、RAD51在GM、ATM+-AT、AT细胞中表达量增高,其中GM、ATM+-AT细胞呈现共定位表达,AT细胞无共定位表达;GM、ATM+-AT细胞中BRCA1、RAD51蛋白表达量与AT细胞比较差异有统计学意义(P<0·01);GM细胞中BRCA1、RAD51蛋白表达量与ATM+-AT细胞比较差异也有统计学意义(P<0·01)。结论GM细胞和ATM+-AT细胞中存在ATM基因,ATM在射线等因素诱导下可以激活其下游基因BRCA1、RAD51,并且使这两种蛋白表达或表达量增加并且相互作用,共同完成辐射损伤后的细胞修复;外源性ATM转入正常GM细胞后,由于外源性的ATM基因不能完全弥补内源性的ATM突变基因的功能,对其下游基因Brca1“部分”磷酸化,表现为BRCA1、RAD51蛋白表达量均较GM细胞低。
Objective To investigate the expression of BRCA1 and RAD51 proteins in GM, ATM + -AT and AT cells before and after 60Coγ-ray irradiation. Methods Immunofluorescence staining and laser scanning confocal microscopy were used to observe the localization and expression of BRCA1 and RAD51 in the cells after GMO, ATM + -AT and AT cells were irradiated with 0 and 10Gy 60Coγ ray. Results Before 60Co γ-ray irradiation, BRCA1 and RAD51 proteins expressed only a small amount in GM, ATM + -AT and AT cells and had no colocalization, and the expression of BRCA1 and RAD51 was the lowest in AT cells. After 10Gy60Coγ ray irradiation, ATM, ATM and -AT, AT cells, GM and ATM + -AT cells showed colocalization, while AT cells had no colocalisation. The expression of BRCA1 and RAD51 in GM and ATM + -AT cells was statistically different from AT cells (P <0.01). The expression of BRCA1 and RAD51 in GM cells was also significantly different from ATM + -AT cells (P <0.01). Conclusions There are ATM genes in GM cells and ATM + -AT cells. ATM can activate its downstream genes BRCA1 and RAD51 under the induction of radiation and increase the expression or expression of these two proteins and interact with them to complete the post-irradiation Cell repair; exogenous ATM into normal GM cells, as exogenous ATM gene can not completely make up for the function of endogenous ATM mutation gene, its downstream gene Brca1 “part” phosphorylation, manifested as BRCA1, RAD51 Protein expression levels were lower than GM cells.