Effective Metabolic Targeting of Human Osteosarcoma Cells In Vitro and in Orthotopic Nude-mouse Models with Recombinant Methioninase
Effective Metabolic Targeting of Human Osteosarcoma Cells In Vitro and in Orthotopic Nude-mouse Models with Recombinant Methioninase
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DOI：
10.21873/anticanres.11887
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发表时间：
2017-09-01
影响因子：
2
通讯作者：
Hoffman, Robert M.
中科院分区：
文献类型：
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作者：
Igarashi, Kentaro;Kawaguchi, Kei;Hoffman, Robert M.
背景：甲硫氨酸依赖可能是唯一已知的癌症的一般代谢缺陷。为了利用甲硫氨酸依赖性进行治疗，我们的实验室先前克隆了L-甲硫氨酸α-脱氨基-γ-巯基甲烷裂解酶[EC 4.4.1.11]）（重组甲硫氨酸酶[rMETase]），随后在各种类型的人类肿瘤的小鼠模型中进行了测试。本研究旨在研究rMETase在体外和体内对人骨肉瘤细胞的疗效。材料与方法：使用WST-8测定法在体外测试人骨肉瘤细胞系143 B、HOS和SOSN 2在暴露于rMET酶72小时期间的存活率。计算体外有效性实验的半数最大抑制浓度。将143 B细胞原位移植到裸鼠胫骨中。在移植后1周将小鼠模型随机分为以下组：第1组，未处理对照;第2组，顺铂（CDDP）[腹膜内（i. p.）注射，每周6 mg/kg，持续3周]，阳性对照;第3组，rMETase，100单位/小鼠，每天腹膜内注射，持续21天。每周一次分别用卡尺和数字天平测量肿瘤大小和体重。结果：rMETase在体外以剂量依赖性方式显著抑制骨肉瘤细胞的生长。在开始后5周，与未处理的对照小鼠相比，CDDP和rMETase处理均显著抑制肿瘤体积。肿瘤体积如下：第1组，未处理，对照：1808.2 +/- 344 mm（3）;第2组，CDDP：1102.2 +/- 316 mm（3），与未处理对照相比p = 0.0008;第3组，rMETase：884.8 +/- 361 mm（3），与未处理对照相比p = 0.0001。所有组中均无动物死亡。小鼠的体重在任何组之间均无显著差异。结论：rMETase对骨肉瘤（一种恶性肿瘤类型）显示出良好的疗效。未来的研究将研究rMETase对骨肉瘤患者源性原位异种移植（PDOX）模型的有效性，作为在临床上测试rMETase的桥梁。
Background: Methionine dependence may be the only known general metabolic defect in cancer. In order to exploit methionine dependence for therapy, our laboratory previously cloned L-methionine alpha-deamino-gamma-mercaptomethane lyase [EC 4.4.1.11]) (recombinant methioninase [rMETase]), which was subsequently tested in mouse models of various types of human tumors. The present study aimed to investigate the efficacy of rMETase on human osteosarcoma cells in vitro and in vivo. Materials and Methods: Human osteosarcoma cell lines 143B, HOS and SOSN2 were tested in vitro for survival during a 72-h exposure to rMETase using the WST-8 assay. Half-maximal inhibitory concentrations were calculated for in vitro efficacy experiments. 143B cells were orthotopically transplanted into the tibia of nude mice. Mouse models were randomized into the following groups 1 week after transplantation: Group 1, untreated control; Group 2, cisplatinum (CDDP) [intraperitoneal (i.p.) injection at 6 mg/kg weekly, for 3 weeks], positive control; Group 3, rMETase, 100 units/mouse i.p. daily, for 21 days. Tumor sizes and body weight were measured with calipers and a digital balance once per week, respectively. Results: rMETase significantly inhibited osteosarcoma cell growth, in a dose-dependent manner, in vitro. Both CDDP and rMETase treatment significantly inhibited tumor volume compared to untreated control mice at 5 weeks after initiation. Tumor volumes were as follows: Group 1, untreated, control: 1808.2 +/- 344 mm(3); Group 2, CDDP: 1102.2 +/- 316 mm(3), p = 0.0008 compared to untreated control; Group 3, rMETase: 884.8 +/- 361 mm(3), p = 0.0001 compared to untreated control. There were no animal deaths in any group. The body weight of mice was not significantly different between any group. Conclusion: rMETase showed promising efficacy against osteosarcoma, a recalcitrant tumor type. Future studies will investigate the efficacy of rMETase on patient-derived orthotopic xenograft (PDOX) models of osteosarcoma as a bridge to testing rMETase in the clinic.