不同给药方式下三氧化二砷对T细胞淋巴瘤EL4细胞体内及体外抑制作用

Effects of arsenic trioxide under different administration ways on T-cell lymphoma xenografts in nude mice:in vivo and in vitro experiments

背景与目的:体外实验证明,三氧化二砷(arsenic trioxide,As2O3)单药可抑制多种恶性淋巴瘤细胞的增殖,并呈时间依赖性和浓度依赖性。临床研究也提示,As2O3单药治疗多种病理亚型淋巴瘤有效。但目前As2O3的剂量、具体用法仍未确定。本实验研究不同给药方式As2O3对鼠源性T细胞淋巴瘤EL4细胞体内外的抗瘤作用,旨在了解As2O3不同给药方法对T细胞淋巴瘤疗效及不良反应。方法:MTT法检测8种浓度As2O3对EL4细胞的抑制作用,流式细胞仪分析、AnnexinⅤ-FITC/PI双标记检测细胞凋亡,电镜观察凋亡形态变化。建立EL4细胞裸鼠移植瘤模型,观察不同方案腹腔注射As2O3对EL4裸鼠移植瘤的抑制作用及裸鼠的耐受情况。结果:与对照组相比,不同浓度As2O3对EL4细胞均存在直接抑制作用(P<0.05),作用72h时的IC50为1.28μmol/L。体内实验显示,4mg·(kg·d)-1×7d与2mg·(kg·d)-1×14d给药对裸鼠移植瘤的抑制作用相近,抑瘤率分别为58.8%和55.6%(P=0.351)。移植瘤组织凋亡细胞增多并可见凋亡小体生成。毒性表现主要为急性肝损害的病理学变化。结论:As2O3体外可抑制EL4细胞增殖并可以诱导细胞凋亡,在总剂量相同的情况下,4mg·(kg·d)-1×7d与2mg·(kg·d)-1×14d的给药方式对裸鼠移植瘤生长的抑制作用近似。

Background and Objective： In vitro, arsenic trioxide （As2O3） can inhibit proliferation of many lymphoma cell lines. In clinic, it also can be used to treat many subtypes of lymphoma. But the dosage and administration ways are undetermined yet. In this research, we studied the antitumor effect of As203 with different administration ways on T-cell lymphoma, and observe the toxicity. Methods： Murine T-cell lymphoma cell line EL4 was treated with As2O3 of 8 concentrations. Cell proliferation was detected by MI-I-assay. Cell apoptosis was evaluated by flow cytometry with Annexin-V-FITC/PI staining, and observed under electroscope and fluorescent microscope. EL4 cells were inoculated into nude mice to establish lymphoma models. The effect of As2O3 on lymphoma in nude mice was observed. Results： As2O3 inhibited the proliferation of EL4 cells with a 50% inhibition concentration （IC50） of 1.28 μmol/L at 72 h （P〈0.05）. When treated with the same total dose of As203 by 4 mg·（kg·d）^-1 for 7 days or 2 mg.（kg.d）-1 for 14 days, the inhibition rates of tumor growth in mice were equivalent （58.8% vs. 55.6%, P=0.351）. Apoptotic cells were increased and apoptotic bodies were observed in xenograft tumor tissues. Acute liver damage is the major toxicity. Conclusion： Shortening the administration course and increasing the daily dosage of As2O3 can be considered as a reasonable model for treating advanced/refractory lymphomas.