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目的检测3种肝细胞系(L02、Hep G2和Hep G2.2.15细胞)中Yes相关蛋白(YAP)和磷酸化Yes相关蛋白(p-YAP)的表达,并探讨经脂多糖(LPS)刺激后对其表达的影响。方法实验选取3种不同肝细胞系(L02、Hep G2和Hep G2.2.15细胞)培养48 h,采用荧光定量PCR检测YAP1/2基因的表达、蛋白印迹(Western blot)检测YAP/p-YAP蛋白的表达;用不同浓度LPS分别刺激上述3种肝细胞系12 h,采用流式细胞术检测LPS刺激对YAP/pYAP表达的影响。结果 YAP在Hep G2和Hep G2.2.15细胞中的表达明显高于其在L02细胞中的表达(P<0.05),且在Hep G2.2.15细胞中增加的幅度更为显著(P<0.05);p-YAP在Hep G2和Hep G2.2.15细胞中表达显著高于其在L02细胞中的表达(P<0.05)。LPS处理3种肝细胞系后,与对照组相比,YAP/p-YAP的表达在L02细胞中明显增加(P<0.05),在Hep G2和Hep G2.2.15细胞中明显降低(P<0.05)。结论 YAP在肝癌细胞中的表达高于正常肝细胞,LPS刺激对L02、Hep G2和Hep G2.2.15 3种肝细胞系中YAP的表达有一定的影响。
Objective To investigate the expression of Yes-associated protein (YAP) and Yes-associated protein (p-YAP) in three hepatocyte lines, L02, Hep G2 and Hep G2.2.15, and to investigate the effect of lipopolysaccharide (LPS) The impact on their expression. Methods Three different hepatocytes (L02, Hep G2 and Hep G2.2.15 cells) were selected and cultured for 48 h. The expression of YAP1 / 2 gene was detected by real-time PCR. The expression of YAP / p-YAP protein was detected by Western blot . The three kinds of hepatocytes were stimulated with LPS at different concentrations for 12 h, respectively. The effect of LPS stimulation on the expression of YAP / pYAP was detected by flow cytometry. Results The expression of YAP in Hep G2 and Hep G2.2.15 cells was significantly higher than that in L02 cells (P <0.05), and more significantly in Hep G2.2.15 cells (P <0.05). The expression of p-YAP in Hep G2 and Hep G2.2.15 cells was significantly higher than that in L02 cells (P <0.05). Compared with the control group, the expression of YAP / p-YAP was significantly increased in L02 cells (P <0.05) and significantly decreased in Hep G2 and Hep G2.2.15 cells (P <0.05) ). Conclusion The expression of YAP in hepatocellular carcinoma cells is higher than that of normal liver cells. LPS stimulated the expression of YAP in L02, Hep G2 and Hep G2.2.15 hepatocytes.