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目的:研究不同剂量的双酚A(BPA)暴露对小鼠精原干细胞C18-4的增殖及表观遗传机制的影响。方法:C18-4细胞暴露于不同浓度的BPA(10-9~10-4 mol/L)96 h,用CCK-8方法测定细胞的增殖;通过点杂交、Real-time PCR、Western blotting方法检测10-5 mol/L BPA、10-9 mol/L BPA对DNA甲基化水平、组蛋白甲基转移酶基因、组蛋白H3K27Me3、H3K36Me3水平的变化。结果:110-9~10-6 mol/L BPA促进C18-4细胞增殖;10-5 mol/L BPA抑制细胞增殖,促进细胞凋亡;10-4 mol/L BPA导致细胞死亡。210-5 mol/L高剂量BPA引起C18-4细胞DNA整体甲基化水平下降(P<0.01)。3 10-5 mol/L高剂量BPA能引起H3K27Me3、H3K36Me3水平下降(P<0.01)。结论:BPA能干扰小鼠精原干细胞C18-4的细胞增殖;高剂量BPA能引起DNA甲基化水平和组蛋白甲基化水平的下降。
Objective: To study the effects of different doses of BPA exposure on the proliferation and epigenetic mechanism of mouse spermatogonial stem cells C18-4. Methods: C18-4 cells were exposed to different concentrations of BPA (10-9-10-4 mol / L) for 96 h. The proliferation of C18-4 cells was detected by CCK-8 assay. Real-time PCR and Western blotting were used to detect the cell proliferation. The levels of DNA methylation, histone methyltransferase gene, histone H3K27Me3, H3K36Me3 in 10-5 mol / L BPA and 10-9 mol / L BPA were measured. Results: 110-9 ~ 10-6 mol / L BPA promoted the proliferation of C18-4 cells; 10-5 mol / L BPA inhibited cell proliferation and promoted cell apoptosis; 10-4 mol / L BPA led to cell death. 210-5 mol / L BPA caused a decrease of DNA methylation in C18-4 cells (P <0.01). 3,10-5 mol / L high dose of BPA can cause H3K27Me3, H3K36Me3 levels decreased (P <0.01). Conclusion: BPA can interfere with the proliferation of C18-4 cells in spermatogonial stem cells of mice. High dose of BPA can induce the decrease of DNA methylation and histone methylation.