氢醌对人L-02肝细胞泛素结合酶Rad6B表达的影响

Effect of hydroquinone on the expression of ubiquitin-conjugating enzyme Rad6B in human L-02 hepatic cells

目的研究氢醌(HQ)对人L-02肝细胞中泛素结合酶Rad6B表达的影响。方法将L-02肝细胞用不同浓度(0、5、10、20、40、80和160μmol/L)的HQ处理24h之后,采用实时荧光定量PCR技术检测Rad6B在mRNA水平上的表达;采用Western blot方法检测Rad6B在蛋白质水平上的表达。结果在0～160μmol/L的范围内,HQ可诱导Rad6B在mRNA和蛋白质水平上表达的增加,具有随着剂量的增加而升高的趋势;并且各染毒剂量组(5、10、20、40、80和160μmol/L组)肝细胞Rad6B在mRNA和蛋白质水平上的表达均明显地高于对照组(0μmol/L组)(P<0.01);当HQ的作用浓度达到160μmol/L时,Rad6B的表达水平达到最大值,其mRNA和蛋白质表达的相对定量值分别为4.35和0.85。结论HQ可使L-02肝细胞的Rad6B表达增加。

Objective To study the effects of hydroquinone(HQ) on expression of ubiquitin-conjugating enzyme Rad6B in human L-02 hepatic cells. Methods After L-02 hepatic cells were at the concentrations of 0,5,10,20,40,80,and 160μmol/L HQ for 24h,and the levels of Rad6B mRNA and protein expressions in L-02 hepatic cells were detected by real-time fluorescent quantitative polymerase chain reaction(QPCR) technique and Western blot method respectively. Results It was found that HQ in the range of 0-160μmol/L could induce increases in the expressions of Rad6B mRNA and protein, which was in a dose-dependent manner. The levels of Rad6B mRNA and protein expressions in the treated group of 5,10,20,40,80,and 160μmol/L were higher than those in the control(P<0.01), levels of Rad6B mRNA and protein expressions reached the maximun when L-02 hepatic cells were treated by HQ at the concentrations of 160μmol/L,and the relative quantity values were 4.35 and 0.85,respectively. Conclusion HQ could regulate up the expression of Rad6B in L-02 hepatic cells.