Laboratory of Molecular Target Therapy for Cancer, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, 465 Kajii-cho, Kawaramachi-Hirokoji, Kamigyo-ku, Kyoto, 602-8566, Japan.
Int J Clin Oncol. 2019 Sep;24(9):1020-1029. doi: 10.1007/s10147-019-01451-9. Epub 2019 Apr 16.
Methotrexate (MTX) is used in first-line treatment of primary central nervous system lymphoma (PCNSL), but most cases result in relapse-acquired resistance to MTX. However, only few studies have reported on internal changes and chemotherapies in PCNSL.
In this study, we generated two MTX-resistant PCNSL cell lines, designated MTX-HKBML and MTX-TK, in addition to a MTX-resistant Burkitt lymphoma cell line, designated MTX-RAJI. We examined gene expression changes and drug sensitivity to a proteasome inhibitor, bortezomib, in these cells.
Cytotoxic tests revealed that the 50% inhibitory concentration for MTX in MTX-HKBML is markedly higher than that in the other two cell lines. Expression of the genes in MTX and folate metabolisms, including gamma-glutamyl hydrolase and dihydrofolate reductase, are upregulated in both MTX-HKBML and MTX-TK, whereas the gene expression of folylpolyglutamate synthetase, thymidylate synthase, and methylenetetrahydrofolate dehydrogenase 1 were upregulated and downregulated in MTX-HKBML and MTX-TK, respectively, on the other hand, bortezomib sensitivity was observed in MTX-TK, as compared with control TK, but not in MTX-HKBML.
These results indicate the cell-type-specific changes downstream of metabolic pathways for MTX and folate, bortezomib sensitivity, and purine and pyrimidine syntheses, in each PCNSL cell line. The MTX-resistant lymphoma cell lines established may be useful for in vitro relapse models for MTX and development of salvage chemotherapy and drug discovery.
甲氨蝶呤(MTX)用于原发性中枢神经系统淋巴瘤(PCNSL)的一线治疗,但大多数病例会对 MTX 产生获得性耐药。然而,仅有少数研究报道过 PCNSL 中的内部变化和化疗。
在这项研究中,我们除了建立甲氨蝶呤耐药的 Burkitt 淋巴瘤细胞系 MTX-RAJI 外,还建立了两个甲氨蝶呤耐药的 PCNSL 细胞系,分别命名为 MTX-HKBML 和 MTX-TK。我们检测了这些细胞中甲氨蝶呤和叶酸代谢相关基因的表达变化以及对蛋白酶体抑制剂硼替佐米的药物敏感性。
细胞毒性试验表明,MTX-HKBML 对 MTX 的 50%抑制浓度明显高于其他两种细胞系。在 MTX-HKBML 和 MTX-TK 中,甲氨蝶呤和叶酸代谢相关基因的表达上调,包括γ-谷氨酰水解酶和二氢叶酸还原酶,而叶酸多聚谷氨酸合成酶、胸苷酸合成酶和亚甲基四氢叶酸脱氢酶 1 的基因表达在 MTX-HKBML 和 MTX-TK 中分别上调和下调。另一方面,与对照 TK 相比,只有在 MTX-TK 中观察到硼替佐米的敏感性,而在 MTX-HKBML 中则没有。
这些结果表明,在每个 PCNSL 细胞系中,存在甲氨蝶呤和叶酸代谢途径下游的细胞类型特异性变化、硼替佐米敏感性、嘌呤和嘧啶合成,这些建立的甲氨蝶呤耐药淋巴瘤细胞系可能有助于 MTX 体外复发模型的建立以及挽救化疗和药物发现的研究。