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目的探讨Toll样受体(Toll-like receptor,TLR)信号转导通路中关键节点蛋白在人肝癌细胞株QGY中的表达及其对QGY细胞增殖能力的影响。方法通过RT-PCR和Western blot检测人正常肝细胞株LO2和QGY细胞株中TLR信号转导通路关键节点mRNA和蛋白的表达水平;采用TRAF-6-siRNA沉默关键节点蛋白TRAF-6的表达,Western blot检测沉默效果,MTT法检测TRAF-6基因沉默后QGY细胞的增殖情况。结果 QGY细胞与LO2细胞相比,MAL和TRAF-6基因表达水平明显升高(P<0.05),TRAM和TRIF基因表达水平差异无统计学意义(P>0.05);转染siRNA后QGY细胞TRAF-6蛋白的表达水平明显降低(P<0.05),细胞存活率明显下降(P<0.05)。结论 TLR信号转导通路中关键节点蛋白MAL和TRAF-6在肝癌细胞中的表达水平高于在正常肝细胞中的表达水平;TRAF-6对肝癌细胞的增殖具有一定的促进作用,有望成为分子靶向治疗原发性肝癌新的作用靶点。
Objective To investigate the expression of key node proteins in Toll-like receptor (TLR) signal transduction pathway in human hepatoma cell line QGY and its effect on the proliferation of QGY cells. Methods RT-PCR and Western blot were used to detect the mRNA and protein expression of key TLR signaling pathway in human normal hepatocyte cell line LO2 and QGY. TRAF-6-siRNA silenced the expression of TRAF-6, The silencing effect was detected by Western blot. The proliferation of QGY cells after TRAF-6 gene silencing was detected by MTT assay. Results Compared with LO2 cells, the expression levels of MAL and TRAF-6 in QGY cells were significantly increased (P <0.05), while there was no significant difference in TRAM and TRIF gene expression between QGY cells and LO2 cells (P> 0.05) -6 protein expression was significantly lower (P <0.05), cell survival rate decreased significantly (P <0.05). Conclusion The expression of MAL and TRAF-6, the key node proteins in TLR signal transduction pathway, is higher than that in normal liver cells. TRAF-6 can promote the proliferation of hepatocellular carcinoma cells and is expected to become a molecule Targeted treatment of primary liver cancer a new target.