MC004对大鼠胚胎中脑细胞增殖和分化的影响  

Effects of MC004 on rat embryo midbrain cell proliferation and differentiation

作　　者：骆永伟[1] 施畅[1] 吴纯启[1] 王全军[1] 柳艳青[1] 杨保华[1] 胡中慧[1] 张玉杰[2] 廖明阳[1] 

机构地区：[1]军事医学科学院毒物药物研究所国家北京药物安全评价研究中心,北京100850 [2]北京美迪克斯生物技术有限公司,北京100012

出　　处：《药物不良反应杂志》2009年第2期96-99,共4页Adverse Drug Reactions Journal

基　　金："重大新药创制科技重大专项"资助项目(2008ZX09305-003);北京市科委科技计划"创新药物研究开发"资助项目(D07080201330721);"北京市自然科学基金"资助项目(7092079)

摘　　要：目的:探讨新型抗肿瘤化合物MC004对胚胎发育的影响。方法:从孕13日龄大鼠胚胎中采集中脑细胞,将其稀释到细胞密度为5×106/ml的细胞悬液,细胞存活率>95%。6孔板和96孔板的每个孔分别注入10μl细胞悬液,在37℃5%二氧化碳培养箱孵化2h。孵化后,6孔板每孔分别加入不同浓度的MC004(5、2.5、1.25、0.625、0.3125、0.15625、0.078、0.039、0.0195、0.0098、0.0049μg/ml)终体积为2ml,96孔板的每个孔分别加入不同浓度的MC004(5、2.5、1.25、0.625、0.3125、0.15625、0.078、0.039、0.0195μg/ml)终体积为200μl,对照组采用相应体积的Ham’sF-12完全培养基,体外培养5d后,弃培养液,细胞固定和染色。在光学显微镜下计数6孔板内微团集落数目,计算细胞分化半数抑制浓度。用酶标仪570nm波长处测定96孔板各孔吸光值,计算细胞增殖半数抑制浓度。结果:不同MC004剂量组细胞集落和集落间神经纤维束减少,有明显量效关系。MC004半数细胞分化抑制浓度为0.0624μg/ml,MC004半数细胞增殖抑制浓度为0.1963μg/ml。结论:MC004在体外能非特异性地抑制细胞增殖和分化,可能具有致畸作用。Objective： To evaluate the effects of the new antineoplastic compound MC004 on embryonic development. Methods： Midbrain cells were harvested from 13-day-old rat embryos, and diluted until the resulting cell density of suspension was 5×10^6/ml; the survival rate of the cells was 〉95%. Each well in 6 and 9 well plates was respectively poured into 10 μl of cell suspension, then was incubated at 37℃ with 5 % carbon dioxide for 2 hours. After incubation, different concentrations of MC004 （5, 2.5, 1.25, 0. 625, 0.312 5,0. 156 25,0. 078, 0.039, 0. 019 5,0. 009 8, 0.0049μg/ml） was added to each well in 6 well plates until the final volume was 2 ml, and different concentrations of MC004（5,2.5, 1.25,0. 625,0. 312 5,0. 156 25,0.078, 0.039, 0. 019 5μg/ml） was added to each well in the 96 well plates until the final volume was 200μl. The corresponding volume of Ham＇ s F-12 complete medium was used in the control group. After 5 days of culture in vitro, the culture medium was removed and the cells were fixed and stained. The number of colony formation in 6 well plates was counted under light microscopy, then the 50% inhibition concentration of differentiation was calculated. The cells absorbency of each well in 96 well plates was measured with micro-plate reader at 570 nm, then the 50% inhibition concentration of proliferation was calculated. Results ： In different MC004 dose groups, the number of colony formation and nerve fiber bundle among colonies decreased. The dose-effect relationship was marked. The 50% inhibition concentration of differentiation to MC004 was 0. 062 4μg/ml. The 50% inhibition concentration of proliferation to MC004 was 0. 196 3 μg/ml. Conclusion： MC004 could nonspecifically inhibit cell proliferation and differentiation in vitro, and it might have teratogenetic effect.

关 键 词：MC004 胚胎中脑细胞 致畸性 细胞增殖 细胞分化 

分 类 号：R341[医药卫生—基础医学]