新型环磷酰胺类衍生物4,6-二苯基环磷酰胺抗肿瘤增殖作用研究

Anti-Tumor Proliferative Effects of a Novel 4,6-Diphenyl Cyclophosphamide

目的研究新型环磷酰胺类衍生物4,6-二苯基环磷酰胺(9b)对体外培养的SK-OV3(人卵巢癌细胞系)、143B(人骨肉瘤细胞系)细胞和体内S180(小鼠肉瘤细胞系)、B16(小鼠黑色素瘤细胞系)实体瘤的抗肿瘤活性,并探讨其可能的机制。方法采用四甲基偶氮唑蓝法观察4,6-二苯基环磷酰胺对SK-OV3和143B细胞增殖的抑制作用;流式细胞仪检测4,6-二苯基环磷酰胺对细胞周期分布和凋亡的影响;体内建立S180、B16荷瘤小鼠模型,观察4,6-二苯基环磷酰胺对瘤体质量的影响。结果四甲基偶氮唑蓝法试验结果显示4,6-二苯基环磷酰胺在3.7~300μmol·L-1和24~72 h内能明显抑制SK-OV3和143 B细胞的增殖,且呈剂量-效应和时间-效应关系;4,6-二苯基环磷酰胺处理SKOV3、143 B细胞48 h的IC50分别为(71.27±1.08)、(98.50±0.82)μmol·L-1;流式细胞结果显示4,6-二苯基环磷酰胺使143B细胞G0/G1期细胞和SK-OV3细胞G0/G1和G2/M期细胞较对照组明显增高(P<0.05,P<0.01),细胞凋亡率明显增加(P<0.05,P<0.01);小鼠S180和B16实体瘤模型中,4,6-二苯基环磷酰胺(3、10和30 mg·kg-1·d-1)对肿瘤的生长也有一定的抑制作用。结论 4,6-二苯基环磷酰胺具有体内外抗肿瘤活性,其抗肿瘤机制可能与细胞周期改变和诱导肿瘤细胞凋亡相关。

ABSTRACT：OBJECTIVE To investigate the anti-tumor activity of a novel cyclophosphamide derivate 4,6-diphenyl cyclophos- phamide （9b） on cultured SK-OV3 （human ovarian cancer cell line） and 143B （human osteosarcoma cell line） cells in vitro and $180 （mouse sarcoma ce]l ]ine） and B16 （murine melanoma cell line） solid tumors in vivo, and its possible anti-tumor mechanism. METHODS The inhibitory effects of 9b on SK-OV3 and 143B cells proliferation were analyzed by MTT assay in vitro. The effect of 9b on cell cycle distribution and apoptosis were evaluated by flow cytometry. To evaluate the anti-tumor effect of 9b in vivo, mouse model bearing inoculated S180 and B16 tumor were established. RESULTS MTT assay revealed that 9b obviously suppressed SK-OV3 and 143B cells growth and proliferation in a dose-dependent manner within the concentration ran- ging from 3.7 to 300 ~mol ~ L-land in a time-dependent manner from 24 to 72 h. The ICs0 value of 9b was （71.27 ＋_ 1.08） txmol ~ L-l for SK-OV3 cells, and （98.50 _＋0.82） p^mol ~ L-1 for 143B cells respectively when incubation for48 h. The re- sults of flow cytometry indicated that after treated for 48 h with different concentration of 9b, the ratios of 143 B cells in the Go/G1 phase and SK-OV3 cells in the G0/G1 phase and G2/M phase were significantly increased compared with control group （ P 〈 0.05, P〈0.01）, the apoptosis rate bad significantly increased （P〈0.05, P〈0.01）. 9b （3, 10 and 30 mg＂ kg-1 ~ d-1） could significantly reduce tumor weight in the S180 and B16 solid tumor mouse model in vivo. CONCLUSION 9b showed sig- nificantly anti-tumor activity both in vivo and in vitro, of which the mechanism might be associated with the change of cell cycle distribution and induction of tumor cell apoptosis.