三氧化二砷抑制小鼠B16黑色素瘤生长作用及其机制

目的　探讨三氧化二砷 (As2 O3 )对小鼠B16黑色素瘤的生长及其血管生成的抑制作用 ;同时观察其对B16细胞增殖活性、细胞形态、细胞周期及凋亡的影响。方法　选用小鼠黑色素瘤B16细胞接种C5 7BL/ 6J小鼠 ,观察腹腔注射As2 O3 对实体瘤的重量及成瘤率的影响 ;应用HE染色、Ⅷ RAg免疫组化染色观测瘤组织内新生血管密度 ;采用CellTiter 96AqueousOne试剂检测B16细胞增殖活力 ;Giem sa染色、Feulgen染色观察细胞形态学变化 ;流式细胞术分析细胞周期及细胞凋亡。结果　As2 O3 能显著抑制小鼠B16黑色素瘤的生长 ,治疗组成瘤率为 37 5 % ,抑瘤率达81 6 1% ,并能显著抑制瘤组织内血管生成 ;体外实验观察到As2 O3 能抑制B16细胞增殖 ,并存在浓度依赖效应 ,IC50 为32 99μmol·L-1;细胞形态学观察结果显示As2 O3 使B16细收稿日期 :2 0 0 4-0 2 -17,修回日期 :2 0 0 4-0 3 -2 8基金项目 :安徽省教育厅资助项目 ,No 2 0 0 4kJ2 79作者简介 :夏　俊 ( 1965 -) ,女 ,硕士 ,副教授 ,硕士生导师 ,研究方向 :肿瘤分子生物学 ,Tel:0 5 5 2 3 0 664 12 2 0 97,E mail:xia jun1965 @yahoo .com .cn崔秀云 ( 1941-) ,女 ,教授 ,博士生导师 ,研究方向 :癌基因与抑癌基因 ,Tel:0 411 472 0 64 8,E mail:cuixy @dlmedu .edu .

Inhibitory effects and mechanism of arsenic trioxide on the growth of melanoma B16 cells

Aim The present study was designed to investigate the growth inhibition and anti-angiogenesis of arsenic trioxide (As 2O 3) on B16 cells xenografts in C57BL/6J mice and observe the effects of As 2O 3 on cell proliferation, cell morphology, cell cycle and apoptosis in vitro. Methods Mice melanoma cell line B16 was transplanted into C57BL/6J mice. The weight of tumor and percent of tumor development were examined after As 2O 3 intraperitoneal administration. HE staining and immunohistochemistry for Ⅷ-R Ag were performed to detect the microvessel density in tumor tissues. CellTiter 96 Aqueous One was used to determine the cell proliferation. Giemsa and Feulgen staining were used to observe the morphological changes of the cells. Cell cycle and apoptosis were analyzed by flow cytometry (FCM). Results As 2O 3 significantly inhibited the tumor growth in vivo, with an inhibitory rate of 81.61%. The microvessel density in tumor tissues was obviously reduced after treated with As 2O 3. There was obviously a concentration-dependent relationship between As 2O 3 and the inhibition of B16 cells proliferation (P<0.01). IC 50 was 32.99 μmol·L -1. Cell morphological results showed that cell density was decreased, contents of DNA were lowed, nuclear area was became smaller, and cells had higher differentiation than that of the control group. FCM demonstrated that As 2O 3 at 20 μmol·L -1 arrest B16 cells in G 0-G 1 phase, but higher dose As 2O 3 at 40 μmol·L -1 might induce apoptosis of B16 cells. Conclusion The results suggest that As 2O 3 can inhibit B16 melanoma growth by means of anti-angiogenesis, cell proliferation inhibition, cell cycle block, and apoptosis.