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）苯磺酸水合物（XTT试剂测定试剂盒）的比色测定法测定细胞生长\u002F活力。在490nm处读取吸光度。通过台盼蓝排除试验测定细胞活力;在治疗24和48小时后手动计数活细胞和非活细胞[3]。",{"key":241,"value":242},"动物实验","大鼠[2]通过注射3×106至1×10 7个细胞或植入约50mg的肿瘤片段，在裸鼠或裸Rowett大鼠（Hsd：RH-Fox1rnu）中皮下或原位生长肿瘤异种移植物。携带肿瘤的动物小鼠用载体对照，Alpelisib（NVP-BYL719）或NVP-BKM120（po，每天）以所示剂量治疗。对于功效研究，当皮下植入的肿瘤达到约200mm 3并且每天50mg \u002F kg用Alpelisib（NVP-BYL719）治疗时，招募携带肿瘤的动物。报告的反应是治疗最后一天肿瘤体积相对于第0天（治疗开始）的百分比变化。小鼠[3]通过吸入异氟烷\u002F空气混合物（2％，1 L \u002F min）麻醉5周龄雄性C57Bl \u002F 6J小鼠，然后肌肉注射1×106小鼠MOS-J骨肉瘤细胞。到胫骨，导致软组织肿瘤迅速增长，伴有继发性骨质侵入。肿瘤在8天后出现在注射部位并导致成骨细胞损伤，再现成骨细胞形式的人骨肉瘤。随机分配三组（每组n = 6）C57B1 \u002F 6J以接受安慰剂或Alpelisib（BYL719）（口服给药，每日12.5-50mg \u002F kg）。肿瘤细胞接种后1天开始预防性治疗。随机分配4组6个C57Bl \u002F 6J接受安慰剂（口服0.5％甲基纤维素和腹腔注射水），Alpelisib（BYL719）（口服50mg \u002F kg \u002F天），异环磷酰胺（腹腔注射30）在第一周期间三次mg \u002F kg），或Alpelisib（BYL719）（每天50mg \u002F kg）和异环磷酰胺（30mg \u002F kg，第一周三次）的组合。",{"key":244,"value":245},"参考文献","[1].\nFuret P, et al. Discovery of NVP-BYL719 a potent and selective phosphatidylinositol-3 kinase alpha inhibitor selected for clinical evaluation. Bioorg Med Chem Lett. 2013 Jul 1;23(13):3741-8.\n[2].\nFritsch C, et al. Characterization of the novel and specific PI3Kα inhibitor NVP-BYL719 and development of the patient stratification strategy for clinical trials. Mol Cancer Ther. 2014 May;13(5):1117-29.\n[3].\nGobin B, et al. BYL719, a new α-specific PI3K inhibitor: single administration and in combination with conventional chemotherapy for the treatment of osteosarcoma. Int J Cancer. 2015 Feb 15;136(4):784-96.\n[4].\nVenot Q, et al. Targeted therapy in patients with PIK3CA-related overgrowth syndrome. Nature. 2018 Jun;558(7711):540-546.\n[5].\nDing J, et al. Inhibition of BTF3 sensitizes luminal breast cancer cells to PI3Kα inhibition through the transcriptional regulation of ERα. 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