| 段秋华,封悦,胡莉钧,汪建林,曾宏伟,孙菲,孟庆红,周梦耘,于静萍.聚腺苷二磷酸核糖聚合酶抑制剂对乳腺癌细胞放射敏感性的影响及其机制研究[J].中华放射医学与防护杂志,2023,43(12):945-953 |
| 聚腺苷二磷酸核糖聚合酶抑制剂对乳腺癌细胞放射敏感性的影响及其机制研究 |
| Effect of poly adenosine diphosphate ribose polymerase inhibitor on the radiosensitivity of breast cancer cells and underlying mechanism |
| 投稿时间：2023-07-06 |
| DOI：10.3760/cma.j.cn112271-20230706-00228 |
| 中文关键词: PARP抑制剂 乳腺癌 放射敏感性 DNA损伤修复 细胞凋亡 |
| 英文关键词:PARP inhibitors Breast cancer Radiosentivity DNA damage repair Apoptosis |
| 基金项目:上海市卫健委卫生行业临床研究专项项目(202340160);常州市卫健委重大科技项目(ZD202017);常州市"十四五"高层次卫生人才培养工程拔尖人才(2022CZBJ070);上海中医药大学科技发展基金(23KFL105) |
| 作者 | 单位 | E-mail | | 段秋华 | 南京医科大学附属常州第二人民医院放疗科, 常州 213003 | | | 封悦 | 上海中医药大学附属曙光医院放疗科, 上海 201203 | | | 胡莉钧 | 南京医科大学附属常州第二人民医院放疗科, 常州 213003 | | | 汪建林 | 南京医科大学附属常州第二人民医院放疗科, 常州 213003 | | | 曾宏伟 | 上海中医药大学附属曙光医院放疗科, 上海 201203 | | | 孙菲 | 南京医科大学附属常州第二人民医院放疗科, 常州 213003 | | | 孟庆红 | 南京医科大学附属常州第二人民医院放疗科, 常州 213003 | | | 周梦耘 | 南京医科大学附属常州第二人民医院放疗科, 常州 213003 | | | 于静萍 | 南京医科大学附属常州第二人民医院放疗科, 常州 213003
上海中医药大学附属曙光医院放疗科, 上海 201203 | yujingping700420@sina.com |
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| 中文摘要: |
| 目的 观察聚腺苷二磷酸核糖聚合酶(PARP)抑制剂尼拉帕利及帕米帕利对乳腺癌MCF-7及MDA-MB-436细胞放射敏感性的影响,并对其机制进行探讨。方法 将乳腺癌MCF-7及MDA-MB-436细胞分为空白对照组、尼拉帕利组、帕米帕利组、单纯照射组、尼拉帕利联合照射组、帕米帕利联合照射组。CCK-8法检测药物对细胞增殖的影响,克隆形成实验检测药物对细胞放射敏感性的影响,流式细胞术分析药物联合照射对细胞周期和凋亡的影响,免疫荧光法检测细胞内γ-H2AX焦点数量的变化,qPCR法及Western blot实验检测细胞中FANCG、Bax、Bcl-2 mRNA及蛋白的表达。结果 尼拉帕利及帕米帕利均能抑制乳腺癌MCF-7及MDA-MB-436细胞的增殖,呈时间-剂量依赖性。随着照射剂量的增加,两种细胞的放射生物学参数D0、Dq、SF2逐渐减小,而放射增敏比SERD0、SERDq逐渐增大。尼拉帕利联合照射组及帕米帕利联合照射组与空白对照组相比,G2/M期比例增加(tMCF-7=41.66、44.08,P<0.05;t436=24.69、18.91,P<0.05); G0/G1期比例减少(tMCF-7=8.67、29.61,P<0.05;t436=26.39、29.12,P<0.05);细胞凋亡率显著增加(tMCF-7=11.17、11.71,P<0.05;t436=42.68、15.89,P<0.05)。与空白对照组相比,MCF-7细胞的单纯照射组、尼拉帕利联合照射组在射线照射后2 h,细胞核内γ-H2AX焦点数量显著增加(t=8.89、21.72,P<0.05);照射后24 h单纯照射组细胞核内γ-H2AX焦点数量基本恢复照射前水平,而尼拉帕利联合照射组细胞核内γ-H2AX焦点数量仍大量存在(t=8.82,P<0.05)。 尼拉帕利联合照射组与空白对照组比较,MCF-7细胞的FANCG、Bcl-2 mRNA表达明显减少(tFANCG=14.07,P<0.05;tBcl-2=29.21,P<0.05);Bax mRNA表达明显增多(t=8.90,P<0.05);与空白对照组相比,尼拉帕利联合照射组MCF-7细胞的FANCG、Bcl-2蛋白水平显著下降(tFANCG=7.09,P<0.05;tBcl-2=10.24,P<0.05);Bax蛋白水平显著升高(t=2.90,P<0.05)。结论 PARP抑制剂尼拉帕利及帕米帕利能增加乳腺癌MCF-7及MDA-MB-436细胞的放射敏感性,其机制可能与下调FA-BRCA通路中FANCG的表达,调节凋亡相关基因,抑制DNA损伤修复有关。 |
| 英文摘要: |
| Objective To study the effects of poly adenosine diphosphate ribose polymerase (PARP) inhibitors niraparib and pamiparib on the radiosensitivity of breast cancer cell lines MCF-7 and MDA-MB-436, and to explore its mechanism.Methods MCF-7 and MDA-MB-436 cells were divided into control group, niraparib group, pamiparib group, radiation group, combination group treated with niraparib and radiation, and combination group treated with pamiparib and radiation, respectively. The effects of drugs on cell proliferation and radiosensitivity were measured by CCK-8 assay and colony formation assay, respectively. The effect of drugs combined with radiation on cell cycle and apoptosis were detected by flow cytometry. Immunofluorescence method was used to detect the changes of γ-H2AX focal number of cells. The expressions of FANCG, Bax and Bcl-2 mRNA and protein were detected by qPCR and Western blot, respectively.Results Both niraparib and pamiparib inhibited the proliferation of breast cancer cells MCF-7 and MDA-MB-436 in a time-dose dependent manner. With the increase of irradiation dose, D0, Dq, SF2 value of MCF-7 and MDA-MB-436 cells decreased, and SERD0 and SERDq value increased. Compared with control group, the percentages of cells in G2/M phase were increased (tMCF-7=41.66, 44.08, P<0.05; t436=24.69, 18.91, P<0.05), the percentage of cells in G0/G1 phase were decreased (tMCF-7=8.67, 29.61, P<0.05; t436=26.39, 29.12, P<0.05), and the cell apoptosis rate was significantly increased (tMCF-7=11.17, 11.71, P<0.05; t436=42.68, 15.89, P<0.05) in the combination group. Compared with control group, the number of γ-H2AX foci of MCF-7 cells in the radiation group and combination group treated with niraparib and radiation increased significantly at 2 h after irradiation (t=8.89, 21.72, P<0.05). At 24 h after irradiation, the number of γ-H2AX foci basically returned to normal level in the radiation group but remained at a higher level in the combination group (t=8.82, P<0.05). Compared with control group, the expressions of FANCG and Bcl-2 mRNA decreased (tFANCG=14.07, P<0.05; tBcl-2=29.21, P<0.05), the expression of Bax mRNA increased (t=8.90, P<0.05), and the expression of FANCG and Bcl-2 proteins decreased (tFANCG=7.09, P<0.05; tBcl-2=10.24, P<0.05), while the expression of Bax protein increased (t=2.90, P<0.05) in the combination group.Conclusions PARP inhibitors niraparib and pamiparib can increase the radiosensitivity of breast cancer MCF-7 and MDA-MB-436 cells probably through down-regulating the expression of FANCG in FA-BRCA pathway, up-regulating apoptosis-related genes and inhibiting DNA damage repair. |
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