Additional file 2 of Constitutively active androgen receptor supports the metastatic phenotype of endocrine-resistant hormone receptor-positive breast cancer

Additional file 1. Supplemental materials and methods: in vitro SUMOylation; RT-PCR; Luciferase reporter assays; PLA, mammosphere studies; scratch assays; transcriptomic data analysis. Table S1. List of primer sequences used for the detection of transcripts. Supplemental figures and figure legends: Figure S1. Elevated levels of SUMO isoforms and HSP27 correlate with high probability of metastasis in ET-treated HR+ BCa patients. Figure S2. HyperSUMO conditions promotes AR SUMOylation and enhances its interaction with Hsp27. Figure S3. SUMO stabilizes AR and reduces its proteasomal degradation. Figure S4. SUMO stimulates basal AR transcriptional activity regardless of the AR-luciferase reporter construct. Figure S5. Concurrent targeting of SUMO-modified and unmodified AR decreases TamR-7 BCa growth in 3D cultures.

附加文件1：补充材料与方法：体外SUMO化修饰（SUMOylation）实验、逆转录聚合酶链式反应（RT-PCR）、荧光素酶报告基因检测（Luciferase reporter assays）、邻近连接测定法（Proximity Ligation Assay，PLA）、乳腺球培养实验、划痕实验、转录组数据分析。表S1：用于转录本检测的引物序列列表。补充图表及图表说明：图S1：在内分泌治疗（ET）后的激素受体阳性（HR+）乳腺癌（BCa）患者中，SUMO亚型与热休克蛋白27（HSP27）的高表达与转移高风险呈正相关。图S2：高SUMO化修饰条件可促进雄激素受体（AR）的SUMO化修饰，并增强其与Hsp27的相互作用。图S3：SUMO化修饰可稳定AR并降低其蛋白酶体依赖性降解。图S4：无论使用何种AR荧光素酶报告基因载体，SUMO化修饰均可增强AR的基础转录活性。图S5：同时靶向SUMO化修饰与未修饰的AR，可在三维培养体系中抑制他莫昔芬耐药型7（TamR-7）乳腺癌细胞的生长。