雄黄主要成分As_(2)S_(2)通过“ANP32A-INHAT-H3乙酰化”轴抑制三阴性乳腺癌细胞增殖、迁移的表观遗传调控机制  

The Epigenetic Regulatory Mechanism of As_(2)S_(2) from Realgar in Inhibiting the Proliferation and Migration of Triple-negative Breast Cancer Cells Through the ANP32A-INHAT-H3 Acetylation Axis

作　　者：孙杰杰 张素峰 王曼曼 李庆林 SUN Jiejie;ZHANG Sufeng;WANG Manman;LI Qinglin(School of Pharmacy,Anhui University of Chinese Medicine,Anhui Hefei 230012,China;School of Integrative Chinese and Western Medicine,Anhui University of Chinese Medicine,Anhui Hefei 230012,China;Key Laboratory of Xin’an Medicine,Ministry of Education,Anhui Hefei 230012,China)

机构地区：[1]安徽中医药大学药学院,安徽合肥230012 [2]安徽中医药大学中西医结合学院,安徽合肥230012 [3]新安医学教育部重点实验室,安徽合肥230012

出　　处：《安徽中医药大学学报》2024年第3期86-92,共7页Journal of Anhui University of Chinese Medicine

基　　金：安徽省自然科学基金项目(2308085QH263);安徽省高等学校自然科学研究重点项目(2022AH050525)。

摘　　要：目的探究雄黄主要成分二硫化二砷(As_(2)S_(2))对三阴性乳腺癌(triple negative breast cancer,TNBC)的作用及表观遗传调控机制。方法采用CCK-8、平板克隆形成和细胞划痕实验探究As_(2)S_(2)对人正常乳腺上皮细胞MCF-10A及TNBC细胞增殖和迁移的影响;4D-label free定量蛋白质组学分析挖掘As_(2)S_(2)抗TNBC的潜在干预靶点酸性核磷蛋白家族成员32A(acidic nuclear phosphoprotein family member 32A,ANP32A);慢病毒感染法构建ANP32A过表达敲低细胞株,探究潜在靶点ANP32A对As_(2)S_(2)抗TNBC作用的影响;蛋白质免疫共沉淀和Western blot实验探究As_(2)S_(2)是否通过ANP32A调控TNBC细胞H3乙酰化。结果As_(2)S_(2)对人正常乳腺上皮细胞MCF-10A影响甚微,但显著抑制TNBC细胞增殖和迁移,且呈剂量依赖性;4D-lable free定量蛋白质组学分析结果显示,促癌因子ANP32A被As_(2)S_(2)显著下调,且ANP32A表达影响As_(2)S_(2)在TNBC中的抗增殖和迁移效果。As_(2)S_(2)能下调ANP32A的蛋白水平,抑制乙酰转移酶抑制剂复合物亚基的招募,增加H3乙酰化水平。结论As_(2)S_(2)通过下调ANP32A蛋白调控TNBC细胞中H3乙酰化,抑制TNBC细胞增殖和迁移。Objective To investigate the effect of As_(2)S_(2),the main component of realgar,on triple-negative breast cancer(TNBC)cells and its epigenetic regulatory mechanism.Methods CCK-8 assay,plate colony formation assay,and wound healing assay were used to explore the effect of As_(2)S_(2) on the proliferation and migration of the normal breast epithelial cell line MCF-10A and TNBC cells;4D-label free quantitative proteomic analysis was used to investigate the potential intervention target acidic nuclear phosphoprotein family member 32A(ANP32A)of As_(2)S_(2) in anti-TNBC therapy;lentivirus transfection was used for the overexpression and knockdown of ANP32A to investigate the role of ANP32A in modulating the anti-TNBC activity of As_(2)S_(2);protein co-immunoprecipitation and Western blot were used to investigate whether As_(2)S_(2) regulated H3 acetylation of TNBC cells through ANP32A.Results As_(2)S_(2) had little effect on the normal breast epithelial cell line MCF-10A,but it significantly inhibitied the proliferation and migration of TNBC cells in a dose-dependent manner.4D-label free quantitative proteomic analysis showed that As_(2)S_(2) significantly downregulated the oncogenic factor ANP32A,and the expression of ANP32A significantly affected the anti-proliferation and ant-migration effects of As_(2)S_(2) in TNBC.As_(2)S_(2) downregulated the protein expression level of ANP32A,inhibited the assembly of complex subunits of acetyltransferase inhibitor,and increased the level of H3 acetylation.Conclusion As_(2)S_(2) inhibits the proliferation and migration of TNBC cells by downregulating the level of ANP32A protein and regulating H3 acetylation in TNBC cells.

关 键 词：雄黄 二硫化二砷 酸性核磷蛋白家族成员32A H3乙酰化 表观遗传 三阴性乳腺癌 

分 类 号：R737.9[医药卫生—肿瘤]