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519和RAMOS模型的肿瘤细胞生长分别有91％,99％和95％的有效抑制作用。此外,在DoHH-2和RAMOS模型中利妥昔单抗增强了苯达莫司汀的抗肿瘤作用,但在Granta 519模型中没有[3]。",{"key":224,"value":225},"细胞实验","苯达莫司汀和美法仑对多发性骨髓瘤（MM）细胞的细胞毒性计算为通过MTS测定法测量细胞存活百分比来抑制细胞活力。简而言之，将细胞（1×10 4 \u002F孔）接种在具有递增浓度的药物的96孔板中，并在孵育24,48,72和96小时后进行分析。为此，向每个孔中加入1μg\u002F mL MTS溶液，在37℃下1小时后，用1N异丙醇和HCl（24：1，体积\u002F体积）溶解深蓝色甲crystals晶体。最后，在96孔板读数器中在490nm处测量吸光度。将细胞存活估计为未处理对照的吸光度百分比，并且每个测试一式三份进行。计算每种药物的抑制浓度50（IC50）和25（IC25），即能够将细胞生长分别降低至未处理对照细胞的50％和25％的量，并且测试并行进行使用等毒浓度的苯达莫司汀和美法仑。相对抗性指数（RRI）表示为8226-LR5的IC50与RPMI-8226细胞的IC50的比值[2]。",{"key":227,"value":228},"动物实验","小鼠[3] CB-17 scid小鼠（DoHH-2，Granta 519）或CB-17 scid-bg小鼠（SuDHL-4，RAMOS）接种1×106（DoHH-2，RAMOS），3×106（ SuDHL-4）或5×106（Granta 519）细胞sc在右胁腹。对于侧腹异种移植物，接种体积为0.2mL，由生长培养基和基质胶中50:50的细胞混合物组成。通过电子卡尺每周两到三次测量肿瘤的长度和宽度并且应用以下等式来估计肿瘤体积：V = L×W2 \u002F 2。允许肿瘤达到约250mm 3，并且将小鼠与治疗组和对照组进行大小匹配（第0天）。对于系统性Granta 519肿瘤模型，在第0天通过尾静脉在0.1mL体积的细胞培养基中注射2×10 6个细胞，并在第14天开始治疗。在整个实验过程中对所有动物进行耳标记和单独监测。通过口服管饲法每天一次在Phosal 50PG：PEG400：乙醇的混合物中施用Navitoclax。在第1天，苯达莫司汀和利妥昔单抗分别以25mg \u002F kg和10mg \u002F kg静脉内给药。在苯达莫司汀和利妥昔单抗之前约2小时给予Navitoclax。所有试验均由每组10只小鼠组成。当肿瘤达到> 2000 mm3的大小或监测到任何窘迫迹象时，小鼠被人道地杀死。窘迫的迹象包括失去走动，呼吸困难或体重减轻>每笼平均体重20％[3]。",{"key":230,"value":231},"参考文献","[1].\nLeoni LM, et al. Bendamustine (Treanda) displays a distinct pattern of cytotoxicity and unique mechanistic features compared with other alkylating agents. Clin Cancer Res. 2008 Jan 1;14(1):309-17.\n[2].\nCives M, et al. Bendamustine overcomes resistance to melphalan in myeloma cell lines by inducing cell death through mitotic catastrophe. Cell Signal. 2013 May;25(5):1108-17.\n[3].\nAckler S, et al. Navitoclax (ABT-263) and bendamustine ± rituximab induce enhanced killing of non-Hodgkin's lymphoma tumours in vivo. Br J Pharmacol. 2012 Oct;167(4):881-91.\n[4].\nMinbo Zang, et al. Effects of chemotherapeutic agent bendamustine for non-hodgkin lymphoma on spermatogenesis in mice. 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