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目的研究Src抑制剂PP2对人肝癌细胞HepG2中Src及Tec(tyrosine kinase expressed in hepatocellular carcino-ma)的作用。方法用免疫细胞化学方法检测Src、Tec在HepG2细胞中的表达;RT-PCR和Western blot方法检测不同浓度PP2处理细胞后,HepG2细胞中Src、Tec mRNA及蛋白表达;并用细胞黏附实验及MTT法检测PP2对细胞黏附及生长的影响。结果Src、Tec在人肝癌细胞HepG2中均呈高表达,用Src抑制剂PP2处理HepG2后,Src、Tec mRNA及蛋白表达明显降低,且细胞黏附能力及生长明显被抑制,并呈剂量依赖性,PP2浓度为40μg/ml时细胞抑制最明显。结论Tec是肝癌信号转导过程中重要的联接和调控分子,与Src存在cross-talk,抑制Tec可能影响肝癌细胞的生长和侵袭等生物学行为。
Objective To investigate the effect of Src inhibitor PP2 on Src and Tec in hepatocellular carcinoma-human HepG2 cells. Methods The expression of Src and Tec in HepG2 cells was detected by immunocytochemistry. The expressions of Src and Tec mRNA and protein in HepG2 cells were detected by RT-PCR and Western blot. The cell adhesion and MTT assay The effect of PP2 on cell adhesion and growth was examined. Results Src and Tec were highly expressed in HepG2 cells. The expression of Src and Tec mRNA and protein were significantly decreased after HepG2 treatment with Src inhibitor PP2, and the cell adhesion and growth were significantly inhibited in a dose-dependent manner. The cell inhibition was the most obvious when PP2 concentration was 40μg / ml. Conclusion Tec is an important junction and regulatory molecule in the signal transduction of hepatocellular carcinoma, which has cross-talk with Src and inhibits the biological behavior of Tec, such as cell growth and invasion.