本文選題：三氯乙酸 + L-細(xì)胞�。� 參考：《環(huán)境與健康雜志》2017年02期

【摘要】：目的探討三氯乙酸(TCA)染毒對(duì)L-02細(xì)胞DNA甲基化轉(zhuǎn)移酶1(DNMT1)蛋白及mRNA表達(dá)的影響。方法取處于對(duì)數(shù)生長(zhǎng)期的正常肝細(xì)胞(L-02細(xì)胞),分別加入含0(對(duì)照)、0.1、0.3、0.9 mmo/L TCA的培養(yǎng)液繼續(xù)培養(yǎng)24、48、72 h,同時(shí),設(shè)DNA甲基化酶抑制劑5-氮雜胞苷(5-aza-d C,5μmol/L)處理組,TCA-re組(0.9 mmol/L TCA處理后換正常培養(yǎng)基繼續(xù)培養(yǎng)24 h)和人肝癌(HepG2)細(xì)胞組作為對(duì)照。檢測(cè)細(xì)胞DNMT1蛋白和mRNA的表達(dá)水平。結(jié)果與對(duì)照組比較,各濃度TCA染毒組及5-aza-d C處理組、TCA-re組L-02細(xì)胞DNMT1 mRNA的表達(dá)水平均較低,而HepG2細(xì)胞組DNMT1 mRNA的表達(dá)水平較高,差異均有統(tǒng)計(jì)學(xué)意義(P0.05)。與相同劑量TCA染毒24 h比較,各濃度TCA染毒48、72 h后L-02細(xì)胞DNMT1 mRNA的表達(dá)水平均較低,除0.1、0.9 mmol/L TCA染毒48 h外,差異均有統(tǒng)計(jì)學(xué)意義(P0.05)。且隨著TCA染毒濃度的升高和染毒時(shí)間的延長(zhǎng),L-02細(xì)胞DNMT1 mRNA的表達(dá)水平均呈下降趨勢(shì)。與0.9 mmol/L TCA染毒組比較,TCA-re組L-02細(xì)胞DNMT1 mRNA的表達(dá)水平較高,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。與對(duì)照組比較,各濃度TCA染毒組及5-aza-d C處理組L-02細(xì)胞DNMT1蛋白的表達(dá)水平均較低,除0.1 mmol/L TCA染毒24、48 h及0.3mmol/L TCA染毒24 h外,差異均有統(tǒng)計(jì)學(xué)意義(P0.05);而TCA-re組L-02細(xì)胞和HepG2細(xì)胞組DNMT1蛋白的表達(dá)水平均無明顯變化。與相同劑量TCA染毒24 h比較,各濃度TCA染毒48、72 h后L-02細(xì)胞DNMT1蛋白的表達(dá)水平均較低,差異均有統(tǒng)計(jì)學(xué)意義(P0.05)。除TCA染毒24 h時(shí)L-02細(xì)胞DNMT1蛋白的表達(dá)水平隨染毒濃度的升高而呈先升高后下降的趨勢(shì)外,隨著TCA染毒濃度的升高和染毒時(shí)間的延長(zhǎng),L-02細(xì)胞DNMT1蛋白的表達(dá)水平均呈下降趨勢(shì)。與0.9 mmol/L TCA染毒組比較,TCA-re組L-02細(xì)胞DNMT1蛋白表達(dá)水平較高,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論 TCA體外染毒可能通過抑制DNMT1的表達(dá)維持或者促進(jìn)細(xì)胞的DNA低甲基化狀態(tài)。
[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.
【作者單位】： 深圳市光明新區(qū)疾病預(yù)防控制中心職業(yè)衛(wèi)生科;深圳市寶安區(qū)疾病預(yù)防控制中心職業(yè)衛(wèi)生科;
【基金】：深圳市寶安區(qū)科技計(jì)劃社會(huì)公益項(xiàng)目(2014264)
【分類號(hào)】：R135.1

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