三氯乙酸对L-02细胞DNA甲基化转移酶1蛋白及mRNA表达的影响

发布时间：2018-06-23 07:25

本文选题：三氯乙酸 + L-细胞　；参考：《环境与健康杂志》2017年02期

【摘要】：目的探讨三氯乙酸(TCA)染毒对L-02细胞DNA甲基化转移酶1(DNMT1)蛋白及mRNA表达的影响。方法取处于对数生长期的正常肝细胞(L-02细胞),分别加入含0(对照)、0.1、0.3、0.9 mmo/L TCA的培养液继续培养24、48、72 h,同时,设DNA甲基化酶抑制剂5-氮杂胞苷(5-aza-d C,5μmol/L)处理组,TCA-re组(0.9 mmol/L TCA处理后换正常培养基继续培养24 h)和人肝癌(HepG2)细胞组作为对照。检测细胞DNMT1蛋白和mRNA的表达水平。结果与对照组比较,各浓度TCA染毒组及5-aza-d C处理组、TCA-re组L-02细胞DNMT1 mRNA的表达水平均较低,而HepG2细胞组DNMT1 mRNA的表达水平较高,差异均有统计学意义(P0.05)。与相同剂量TCA染毒24 h比较,各浓度TCA染毒48、72 h后L-02细胞DNMT1 mRNA的表达水平均较低,除0.1、0.9 mmol/L TCA染毒48 h外,差异均有统计学意义(P0.05)。且随着TCA染毒浓度的升高和染毒时间的延长,L-02细胞DNMT1 mRNA的表达水平均呈下降趋势。与0.9 mmol/L TCA染毒组比较,TCA-re组L-02细胞DNMT1 mRNA的表达水平较高,差异有统计学意义(P0.05)。与对照组比较,各浓度TCA染毒组及5-aza-d C处理组L-02细胞DNMT1蛋白的表达水平均较低,除0.1 mmol/L TCA染毒24、48 h及0.3mmol/L TCA染毒24 h外,差异均有统计学意义(P0.05);而TCA-re组L-02细胞和HepG2细胞组DNMT1蛋白的表达水平均无明显变化。与相同剂量TCA染毒24 h比较,各浓度TCA染毒48、72 h后L-02细胞DNMT1蛋白的表达水平均较低,差异均有统计学意义(P0.05)。除TCA染毒24 h时L-02细胞DNMT1蛋白的表达水平随染毒浓度的升高而呈先升高后下降的趋势外,随着TCA染毒浓度的升高和染毒时间的延长,L-02细胞DNMT1蛋白的表达水平均呈下降趋势。与0.9 mmol/L TCA染毒组比较,TCA-re组L-02细胞DNMT1蛋白表达水平较高,差异有统计学意义(P0.05)。结论 TCA体外染毒可能通过抑制DNMT1的表达维持或者促进细胞的DNA低甲基化状态。
[Abstract]:Objective to investigate the effect of trichloroacetic acid (TCA) on the expression of DNA methyltransferase 1 (DNMT1) protein and mRNA in L-02 cells. Methods normal hepatocytes (L-02 cells) in logarithmic growth phase were cultured in 0 (control) 0.3mmol / L TCA medium for 24 h, 48 h, respectively. TCA-re (0.9 mmol / L TCA) treated with 5-azacytidine (5-aza-d Con 5 渭 mol / L) and human hepatocellular carcinoma (HepG2) cells were cultured in normal medium for 24 h. The expression of DNMT1 protein and mRNA was detected. Results compared with the control group, the expression of DNMT1 mRNA in L-02 cells was lower in TCA-exposed group and TCA-re group, but higher in HepG2 cell group (P0.05). Compared with the same dose of TCA for 24 h, the expression of DNMT1 mRNA in L-02 cells was significantly lower than that in the same dose of TCA for 48 h (P0.05), except 0.1mmol / L TCA for 48 h. The expression of DNMT1 mRNA decreased with the increase of TCA concentration and the prolongation of TCA exposure time. Compared with 0.9 mmol / L TCA group, the expression of DNMT1 mRNA in L-02 cells of TCA-re group was higher than that of 0.9 mmol / L TCA group (P0.05). Compared with the control group, the expression of DNMT1 protein in L-02 cells was lower in TCA-exposed group and 5-aza-d C group, except 0.1 mmol / L TCA for 24 h and 0.3 mmol / L TCA for 24 h. The expression of DNMT1 in L-02 cells and HepG2 cells in TCA-re group showed no significant difference (P0.05). Compared with the same dose of TCA for 24 h, the expression of DNMT1 protein in L-02 cells was significantly lower than that in the same dose of TCA for 4872 h (P0.05). The expression of DNMT1 protein in L-02 cells increased first and then decreased with the increase of TCA concentration 24 h after TCA exposure, and the expression level of DNMT1 protein decreased with the increase of TCA concentration and the prolongation of TCA exposure time. Compared with 0.9 mmol / L TCA group, the expression of DNMT1 protein in L-02 cells of TCA-re group was higher than that of 0.9 mmol / L TCA group (P0.05). Conclusion TCA exposure in vitro may maintain or promote the hypomethylation of DNA by inhibiting the expression of DNMT1.
【作者单位】：深圳市光明新区疾病预防控制中心职业卫生科;深圳市宝安区疾病预防控制中心职业卫生科;
【基金】：深圳市宝安区科技计划社会公益项目(2014264)
【分类号】：R135.1

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