氯化锰对脉络丛Z310细胞增殖及细胞周期的影响

Effect of manganese chloride on cell proliferation and cell cycle in Z310 cells

目的探讨氯化锰(MnCl2)对永生化的脉络丛上皮细胞系Z310细胞增殖作用及细胞周期的影响。方法采用MTT法检测低剂量MnCl2(0.00005、0.0005、0.005、0.05、0.5、5、20和50μmol/L)和高剂量MnCl2(50、100、200、400、800、1000和1600μmol/L)对Z310细胞增殖的影响,同时在光学显微镜下观察MnCl2作用后细胞的形态变化,在透射电镜下观察细胞超微结构的变化,并采用流式细胞术检测不同剂量MnCl2对细胞周期时相的影响。结果在一定范围内的低剂量MnCl2对Z310细胞增殖起促进作用,随着剂量的增高,MnCl2对细胞增殖的影响转变为抑制作用。200、400及800μmol/L的MnCl2作用48 h后,Z310细胞与对照组相比细胞数量明显减少,排列不紧凑,且多边形态不明显,呈类梭状生长,核膜皱缩,核质浓缩、边缘化,线粒体出现缺嵴空泡化,胞质空泡化。染毒组与对照组相比,100、200、400及800μmol/L的MnCl2作用于Z310细胞24 h后,处于G0/G1期的细胞比例较对照组从1.8%增加到7.6%,48 h后从5.0%增加到19.2%。结论低剂量(0.00005～5μmol/L)和高剂量(100～1600μmol/L)MnCl2对脉络丛上皮细胞Z310细胞增殖分别具有低剂量促进和高剂量抑制的作用。高剂量MnCl2能将Z310细胞周期阻滞在G0/G1期,且具有剂量-效应关系,同时高剂量MnCl2还可造成Z310细胞形态及超微结构发生病理性改变。

Objective The study aims to explore the toxicological effect of manganese chloride（ MnC12 ） on cell proliferation and cell cycle in choroidal epithelial Z310 cells. Methods MTT assay was conducted followed by exposure of low dose MnCI2 （0. 00005,0. 0005, O. 005,0.05,0.5,5,20,50 μmol/L） and high dose MnCl2 （50, 100, 200, 400, 800, 1000, 1600 μmol/L） on immortalized epithelial Z310 cells to observe the effect of MnCl2 induced cell proliferation. Morphological changes in cellular and ultra structure level following MnC12exposure were observed under optical and electronic microscope. Cell cycle progression aheration was measured quantitatively by Flow cytometer. Results The results showed that cell proliferation could be stimulated by low dosage of MnC12 and the stimulation effect was strong originally, then weaker, and finally transformed into inhibition effect, as MnCl2 dosage increased gradually. The total amount of cells significantly decreased after 24 h MnCl2 exposure, cell density became less compact than the control group and ceils＇ polygonal phenotypechanged into shuttle-shape. Concentrated nucleuses and crimpled laryotheca could be found under electronic microscope. Compared with control group, G0/G1 phase of Z310 cells exposed to MnC1224h and 48 h with dosage of 50 to 800 μmol/L was arrested significantly, the cell count increased from 1.8% to 7.6% and 5.0% to 19.2% respectively. Conclusion All those results indicate that MnC12 could suppress Z310 cell proliferation and induce GO/G1 phase of Z310 cells arrest. The associated molecular mechanism deserves further investigation.