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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 investigate the effect of a novel anti-tumor compound MC004 on embryonic development. Methods: The midbrain cells were collected from pregnant 13 day old rat embryos and diluted to a cell suspension with cell density of 5 × 106 / ml. The cell survival rate was> 95%. Each well of a 6-well plate and a 96-well plate was filled with 10 μl of the cell suspension and incubated for 2 hours at 37 ° C in a 5% carbon dioxide incubator. After incubation, 6-well plates were added to each well with different concentrations of MC004 (5,2.5,1.25,0.625,0.3125,0.15625,0.078,0.039,0.0195,0.0098,0.0049μg / ml) final volume of 2ml, 96-well plates Each well was added with different concentrations of MC004 (5,2.5,1.25,0.625,0.3125,0.15625,0.078,0.039,0.0195μg / ml) final volume of 200μl, the control group with the appropriate volume of Ham’s F-12 complete medium, After cultured for 5 days in vitro, the culture medium was discarded and cells were fixed and stained. Count the number of micellar colonies in a 6-well plate under an optical microscope and calculate the half-value concentration of cell differentiation. With microplate reader 570nm wavelength determination of 96 well plate absorbance of each well to calculate the cell proliferation half inhibitory concentration. Results: There was a significant dose-effect relationship between the number of colony and colony of CNF in different MC004 dose groups. MC004 half cell differentiation inhibitory concentration of 0.0624μg / ml, MC004 half cell proliferation inhibitory concentration of 0.1963μg / ml. Conclusion: MC004 can inhibit cell proliferation and differentiation non-specifically in vitro and may have teratogenic effects.