【摘要】：目的探討放射線誘導(dǎo)的乳腺癌1號基因(BRCA1)出核效應(yīng)對同源重組介導(dǎo)的DNA雙鏈斷裂損傷(DSB)修復(fù)功能的影響及對聚腺苷二磷酸核糖聚合酶(PARP)抑制劑的增敏作用。方法采用亞細(xì)胞純化及蛋白印跡法檢測乳腺癌細(xì)胞株MCF7經(jīng)放射線處理后BRCA1的亞細(xì)胞定位,同時采用免疫熒光法檢測細(xì)胞核磷酸化的H2AX組蛋白(γ-H2AX)、Rad51核焦點形成。流式細(xì)胞技術(shù)檢測細(xì)胞凋亡、克隆形成實驗檢測體外細(xì)胞存活情況,并制作裸鼠皮下移植瘤模型,分為4組,分別為:對照組、放射+DMSO組、假照+ABT-888(ABT-888為PARP抑制劑)組、放射+ABT-888組,每5 d為1個治療周期,周期第1天給予2 Gy照射或假照射。第2~5天給予溶劑(對照組、放射+DMSO組)或20 mg/(kg·d)的ABT-888(假照+ABT-888組),總共給予4個周期的治療。檢測各組的體內(nèi)抑瘤作用。結(jié)果經(jīng)4 Gy放射處理后,MCF7胞核BRCA1表達(dá)量減少,僅為對照組的41%,但胞漿內(nèi)表達(dá)量明顯增多,是對照組的2.14倍(P0.01);同時MCF7細(xì)胞經(jīng)4 Gy放射處理后ABT-888誘導(dǎo)的Rad51核焦點形成陽性細(xì)胞也較對照組明顯減少(7%vs.30%,P=0.01)。放射線和ABT-888聯(lián)合應(yīng)用導(dǎo)致MCF7細(xì)胞凋亡率增高(19%),與對照組(5%)、放射+DMSO組(9%)或假照+ABT-888組(6.2%)相比較差異有統(tǒng)計學(xué)意義(P0.01)。裸鼠移植瘤模型中放射+ABT-888組對腫瘤生長抑制作用明顯優(yōu)于假照+DMSO組或假照+ABT-888組(P0.01)。結(jié)論放射線可誘導(dǎo)BRCA1出核并增強PARP抑制劑的抗腫瘤作用。
[Abstract]:Objective to investigate the effect of radiation-induced nucleation of breast cancer gene 1 (BRCA1) on the repair function of (DSB) induced by homologous recombination of DNA double strand break and to enhance the sensitivity of polyadenosine diphosphate ribose polymerase (PARP) inhibitor. Methods the subcellular localization of BRCA1 in breast cancer cell line MCF7 was detected by subcellular purification and Western blotting. Meanwhile, nuclear phosphorylation of H2AX histone (緯-H2AX) and nuclear focus formation of Rad51 were detected by immunofluorescence. Flow cytometry was used to detect cell apoptosis and clone formation assay was used to detect cell survival in vitro. The nude mice were divided into 4 groups: control group, radiation DMSO group, pseudoradiographic ABT-888 (ABT-888 as PARP inhibitor) group. ABT-888 group was given 2 Gy irradiation or false irradiation every 5 days. A total of 4 cycles of treatment were given on day 2 to day 5: solvent (control group, radiation DMSO group) or 20 mg/ (kg d) ABT-888 (pseudoradiographic ABT-888 group). The inhibitory effect in vivo of each group was detected. Results after 4 Gy irradiation, the expression of BRCA1 in the nucleus of MCF7 was decreased to 41% of that in the control group, but the expression of BRCA1 in the cytoplasm was significantly increased. At the same time, the number of Rad51 focus forming positive cells induced by ABT-888 in MCF7 cells after 4 Gy irradiation was significantly lower than that in the control group (7vs.30). The apoptosis rate of MCF7 cells was increased by radiation combined with ABT-888 (19%), which was significantly higher than that of control group (5%), radiation DMSO group (9%) or pseudoradiographic ABT-888 group (6.2%) (P0.01). In nude mice, the inhibitory effect of radiation ABT-888 on tumor growth was significantly better than that of DMSO or ABT-888 (P0.01). Conclusion radiation can induce the nucleation of BRCA1 and enhance the anti-tumor effect of PARP inhibitor.
【作者單位】： 廣州醫(yī)科大學(xué)附屬第一醫(yī)院呼吸疾病國家重點實驗室 廣州呼吸疾病研究所;廣州醫(yī)科大學(xué)附屬第一醫(yī)院病理科;
【基金】：國家自然科學(xué)基金資助項目(編號:81272901) 廣州市醫(yī)藥衛(wèi)生科技項目(編號:20151A011067)
【分類號】：R737.9

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