Notch1和HES1在骨肉瘤组织中的表达及其对U2OS细胞侵袭能力的影响

Expressions of Notch1 and HES1 in osteosarcoma tissue and their effects on invasion ability of osteosarcoma cell U2OS

目的:研究Notch1和HES1基因在骨肉瘤组织及骨肉瘤细胞系U2OS中的表达及其对U2OS侵袭能力的影响。方法:组织学水平研究中,配对选取23例骨肉瘤及临近瘤旁正常骨骼肌组织标本;细胞学水平研究中,选取骨肉瘤细胞系U2OS及人正常成骨细胞系h FOB1.19;采用免疫组化法检测组织学水平Notch1及HES1蛋白的表达,采用Western blot及Real-time PCR法检测组织学及细胞学水平Notch1、HES1蛋白及mRNA的表达。用Notch1-siRNA沉默U2OS细胞中Notch1基因后,应用Western blot及Real-time PCR法检测U2OS细胞中Notch1及HES1的表达,Transwell法检测U2OS侵袭能力的变化。结果:相比于正常组织,骨肉瘤组织中Notch1、HES1 mRNA和蛋白呈高表达(t配对=22.567,14.830和8.439,7.100,P<0.001)。U2OS细胞中Notch1、HES1 mRNA和蛋白表达强于h FOB1.19细胞(t=11.053,20.482和28.845,34.681,P<0.001)。沉默Notch1基因后,U2OS细胞Notch1、HES1mRNA和蛋白表达下降(t=6.294,9.209和9.151,13.953,P<0.05),侵袭能力下降(t=71.387,P<0.001)。结论:Notch1和HES1在骨肉瘤组织及骨肉瘤细胞系中呈高表达,沉默Notch1基因可降低U2OS细胞侵袭能力。

Aim：To investigate the expressions of Notch 1 and HES1 gene in osteosarcoma tissue and their effects on invasion ability of osteosarcoma cell U 2OS.Methods：Twenty-three matched tissue specimens of osteosarcoma and matched para-tumor normal tissue, and osteosarcoma cell line U2OS and human osteoblast cell line hFOB1.19 were chosen.The ex-pressions of Notch1 and HES1 mRNA and protein in tissue specimens were detected by immunohistochemistry ,Real-time PCR and Western blot.The expressions of Notch1 and HES1 mRNA and protein in hFOB1.19,U2OS, and U2OS transfect-ed with Notch1-siRNA were detected by Real-time PCR and Western blot , and the cell invasion ability was detected by Tr-answell chambers .Results：Compared with para-tumor normal tissue , the Notch1 and HES1 mRNA and protein expressions in osteosarcoma tissue were higher（tmatch =22.567,14.830 and 8.439,7.100,P〈0.001）, and those in U2OS were higher than in hFOB1.19（t=11.053,20.482 and 28.845,34.681,P〈0.001）.Compared with U2OS, the Notch1 and HES1 mRNA and protein expressions in U2OS transfected with Notch1-siRNA were lower（t=6.294,9.209 and 9.151,13.953, P〈0.05）, and the invasion ability decreased（t=71.387,P〈0.001）.Conclusion： Notch1 and HES1 gene show high expression in both tissue and cellular level in osteosarcoma;Notch1 silence can reduce the invasion ability of U 2OS.