氟对人胚肝细胞P53表达的影响

Effect of Fluoride on Expression of P53 in Human Embryo Hepatocytes

[目的]探讨氟对人胚肝细胞(L-02细胞)P53表达的影响。[方法]对体外培养的人胚肝细胞染毒氟化钠40、80、160μg/ml24h后,分别采用MTT试验、蛋白印迹技术(Westernblot)和逆转录-聚合酶链式反应(RT-PCR)方法检测人胚肝细胞存活率、P53蛋白和mRNA水平。[结果]高浓度氟组细胞存活率显著低于对照组(P<0.01);中、高浓度氟组细胞P53蛋白水平均显著高于对照组(P<0.01),且随氟浓度增加而升高;低浓度氟组与对照组差异没有显著性(P>0.05);各氟染毒组P53mRNA的表达量均显著高于对照组(P<0.01),低、中浓度氟组P53mRNA表达量随染氟浓度的增高而增加;高浓度氟组的P53mRNA表达量虽然与对照组有显著性差异(P<0.01),但相对于中浓度氟组来说,已呈现出下降的趋势,与低浓度氟组水平相当。[结论]高氟可使人胚肝细胞的存活率下降,并诱导P53表达,但P53mRNA和P53蛋白的变化趋势并不完全一致。

[ Objective ] To investigate P53 expression induced by fluoride in human embryo hepatocytes. [ Methods ] After L-02 cells were incubated with 40, 80, and 160μg/ml sodium fluoride for 24 hours, the cell survival rate, the level of P53 protein and mRNA expression was assessed by MTT, Western blot and RT-PCR analysis respectively. [ Results ] The cell survival rate in L-02 cells treated by high concentration of fluoride was significantly lower than that in the control group. The expression levels of P53 protein in middle and high concentration fluoride group were obviously higher than that in the control group, and increased as the concentration of fluoride increased. Although no significant difference was seen between the low concentration fluoride group and the control group, there was a rising tendency. The P53 mRNA levels in all fluoride groups were markedly higher than that of the control group. In the low concentration group and the medium concentration group, the levels of P53 mRNA were increased with the increasing concentration of fluoride . The P53 mRNA expression of the high concentration fluoride group was statistically different from that of the control group, but compared with the medium concentration group, it seemed to go down, and it＇s similar to the low concentration fluoride group. [ Conclusion ] Fluoride can decrease the cell survival rate and induce P53 expression in L-02 cells, but there was no consistent tendency between the expression of P53 mRNA and protein.