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通路\n>>\n岩石\n信号通路\n>>\nTGF-beta\u002FSmad 信号通路\n>>\n岩石\n研究领域\n>>\n癌症\n天然产物\n>>\n生物碱",{"key":235,"value":236},"靶点","ROCK",{"key":238,"value":239},"体外研究","Narciclasine激活多形性胶质母细胞瘤细胞中的Rho和应力纤维。对6种人类多形性胶质母细胞瘤(U373,Hs683,GL19,GL5,GL16,GL17)计算的平均IC50为~50 nM,一组60个癌细胞系中Narciclasine的平均IC50值为47 nM [1]。伯氏藻提取物的生物测定引导的分级分离导致识别作为生物活性化合物的lycorine。对于Narciclasine,测量胚根生长抑制的IC50为0.1μM[2]。",{"key":241,"value":242},"体内研究","1mg\u002Fkg的Narciclasine静脉注射方案显着(P = 0.02)增加了GL19多形性胶质母细胞瘤小鼠的存活率。 Narciclasine以相同剂量口服给药,每周五次,连续5周也显着增加该模型中的动物存活率(P = 0.008)。使用1 mg\u002Fkg的Narciclasine口服治疗可显着提高Hs683多形性胶质母细胞瘤小鼠的存活率(P = 0.004)。增加每周施用的剂量数不会增加这些Hs683多形性胶质母细胞瘤小鼠的存活率。 Narciclasine似乎在这些模型中显示出与替莫唑胺相似的存活率增加,但是在口服和静脉注射后均可显着降低剂量[1]。",{"key":244,"value":245},"激酶实验","使用RhoA G-LISA测定法。该测定使用涂有Rho家族效应蛋白的Rho结合结构域的96孔板。 Rho家族蛋白的活性GTP结合形式但不是来自生物样品的无活性GDP结合形式将与平板结合。然后通过与特异性一抗孵育，然后与缀合辣根过氧化物酶的二抗孵育，检测结合的活性Rho家族蛋白。然后使用OPD开发信号。 U373和GL19多形性胶质母细胞瘤血清饥饿24小时，未经处理或用Narciclasine（100 nM）处理3分钟，5分钟，30分钟，1小时和2小时，或用2.0μg\u002F mL细胞处理可渗透的Rho抑制剂在37℃的无血清培养基中培养4小时，有或没有随后的Narciclasine处理。该产物由高度纯化的C3转移酶组成，该转移酶通过二硫键与专有的细胞穿透部分共价连接。细胞穿透部分允许通过质膜快速有效地转运。一旦进入胞质溶胶，细胞穿透部分被释放，从而允许C3转移酶在细胞内自由扩散并且无活性的RhoA，RhoB和RhoC扩散但不相关的GTP酶如Cdc42或Rac1。 C3转移酶通过在GTP酶的效应子结合结构域中的Asn41上的ADP核糖基化来抑制Rho蛋白。然后裂解细胞并通过RhoA G-LISA活化测定法测量RhoA活性，或者对细胞进行F-肌动蛋白染色[1]。",{"key":247,"value":248},"细胞实验","细胞[1]用MTT试验评估Narciclasine IC50浓度，即将给定细胞群体的全球生长速率降低50％的Narciclasine浓度。在存在和不存在Narciclasine（浓度范围在10-9和10-5M浓度范围内）的情况下将细胞温育72小时，以测定Narciclasine IC50值[1]。",{"key":250,"value":251},"动物实验","小鼠[1]将Hs683细胞系和GL19原始培养物移植到裸免疫缺陷小鼠的大脑中，均产生侵入性脑肿瘤。携带异种移植物的小鼠单独接受载体，口服替莫唑胺40mg \u002F kg（每周5次给药，连续5周），或口服1mg \u002F kg的Narciclasine（每周一次，持续5周）或静脉注射（每周两次， 5周）。在Hs683和GL19模型的肿瘤移植后第5天和第7天分别开始给药。基于先前优化的方案选择替莫唑胺剂量和治疗方案。基于我们最近公布的口服给药和药代动力学研究后大鼠的Narciclasine毒性研究，选择Narciclasine剂量和治疗方案。在毒性研究中，Narciclasine（25,10或1 mg \u002F kg）每周给药5次，持续3周，无不良反应水平剂量定义为1 mg \u002F kg \u002F d po，最小的棘皮反应变化和在该剂量水平观察到的一些生物化学参数的微小变化被认为是不相反的。",{"key":253,"value":254},"参考文献","[1].\nLefranc F, et al. Narciclasine, a plant growth modulator, activates Rho and stress fibers in glioblastoma cells. Mol Cancer Ther. 2009 Jul;8(7):1739-50.\n[2].\nWahyuni DS, et al. The use of bio-guided fractionation to explore the use of leftover biomass in Dutch flower bulb production as allelochemicals against weeds. 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