Erastin

方法：为检测体内抗肿瘤活性，将 Erastin (20 mg/kg in 20 μL DMSO plus 130 μL corn oil) 腹腔注射给携带人前列腺癌肿瘤 DU145、ARCaP、PC3 或 H660 的 NSG 小鼠，每天一次，持续二至五周。 结果：Erastin 治疗显著抑制人前列腺癌肿瘤的生长，表明在体内具有抗肿瘤活性。[4] 方法：为研究 Erastin 治疗对抗癌辐射效率的影响，将 Erastin (15 mg/kg in 5% DMSO/corn oil) 腹腔注射给携带人肺腺癌肿瘤 NCI-H1975 的 BALB/c Slc-nu/nu 小鼠，每天一次，持续三天。在最后一次注射 Erastin 后 24 h，用 3 Gy 的 X 射线局部照射麻醉的小鼠。 结果：Erastin 治疗的 NCI-H1975 细胞移植小鼠显示出对 X 射线照射的增敏趋势，同时肿瘤内谷胱甘肽浓度降低。[5]

方法：人胃癌细胞 HGC-27 用 Erastin (1-50 μM) 处理 24 h，使用 CCK-8 方法检测细胞生长抑制情况。 结果：Erastin 剂量依赖性地抑制 HGC-27 细胞生长，IC50 约为 14.39 μM。[1] 方法：人黑色素瘤细胞 A375 用 Erastin (2-10 μM) 处理 3-12 h，使用 Western Blot 方法检测靶点蛋白表达水平。 结果：Erastin 处理引起 A375 细胞中 VDAC2 和 VDAC3 的水平显著下调，VDAC1 的水平略有降低。[2] 方法：人结肠癌细胞 HT-29 用 Erastin (0.1-30 μM) 处理 12 h，使用 Flow Cytometry 方法检测细胞内 ROS 水平。 结果：Erastin 处理显著增加了 HT-29 细胞中的 ROS水平。[3]

Ethanol:< 1 mg/mL (insoluble or slightly soluble); DMSO:25 mg/mL (45.7 mM); H2O:< 1 mg/mL (insoluble or slightly soluble)

BJeLR cells were plated at 100,000 cells/dish in 35 mm tissue culture dishes. After 12h cells were treated with vehicle (DMSO; 10 hrs), erastin (37 μM; 10 hrs), staurosporine (750 nM; 8 hrs), hydrogen peroxide (16 mM; 1 hr) or rapamycin (100 nM; 24 hrs). Cells were fixed with 2.5% glutaraldehyde in 0.1 M Sorenson's buffer (0.1 M H2PO4, 0.1 M HPO4 (pH 7.2)) for at least 1 h, and then treated with 1% OsO4 in 0.1 M Sorenson's buffer for 1 h. Enblock staining used 1% tannic acid. After dehydration through an ethanol series, cells were embedded in Lx-112 and Embed-812 (EMS). Thin sections were cut on an MT-7000 ultramicrotome, stained with 1% uranyl acetate and 0.4% lead citrate, and examined under a Jeol JEM-1200 EXII electron microscope. Pictures were taken on an ORCA-HR digital camera at 5,000-50,000-fold magnification [1].

Tumor growth studies were performed in severe combined immunodeficient (SCID) mice xenograft model. Briefly, 2×10^6 viable HT-29 cells in 100 μL of growth medium (per mouse) were subcutaneously inoculated, and mice bearing ~100 mm3 tumors were randomly divided into three groups with 10 mice per group. Mice were treated daily with 10 or 30 mg/kg body weight of erastin (intraperitoneal injection, for 4 weeks) or vehicle control (Saline). Tumor volumes were calculated by the modified ellipsoid formula: (π / 6) ×AB2, where A is the longest and B is the shortest perpendicular axis of a tumor mass. Mice body weights were also recorded every week. Humane endpoints were always utilized to minimize mice suffering. Animals were observed on daily bases. Signs such as significant-reduced locomotion, severe diarrhea, severe piloerection or a sudden weight loss (> 20%) were recorded. If animals reached these endpoints they were euthanized by exsanguination under 2,2,2-tribromoethanol anesthesia (4 mg/10 g body weight). All injections were performed under the 2,2,2-tribromoethanol anesthesia method [3].