摘要
目的:初步探讨活化Notch1信号影响骨肉瘤细胞侵袭行为的分子机制。分析Notch1信号活化后,RANKL-OPG-RANK信号轴的变化情况。方法:体外培养骨肉瘤细胞系Saos-2,转染Notch1信号的胞内段ICN1后,检测骨肉瘤细胞的侵袭能力,利用实时定量PCR检测RANKL、RANK和OPG的mRNA表达水平。结果:实验组(Saos-2-ICN)ICN1的蛋白表达水平高于对照组(Saos-2-G418)。Saos-2-G418侵袭细胞数为(41.4±6.4)个,Saos-2-ICN侵袭细胞数为(93.7±12.8)个。Saos-2-G418组RANKL、OPG和RANK分子mRNA相对表达水平分别为(0.6±0.07),(1.2±0.14)和(0.3±0.04);Saos-2-ICN组RANKL、OPG和RANK分子mRNA相对表达水平分别为(1.8±0.3),(0.58±0.09)和(2.0±0.4),两者间具有统计学差异(P<0.05)。结论:活化Notch1信号,提高细胞侵袭力,升高RANKL和RANK基因mRNA表达水平,降低OPG基因mRNA表达水平。本实验结果提示RANKL-OPG-RANK信号轴可能参与Notch1信号调控骨肉瘤细胞的侵袭和转移。
Objective; To analyze the mechanism for which Notchl activation enhances human osteosarcoma cell invasion ability and to explore RANKL-OPG-RANK signaling axis changes after the Notchl signal activation, Methods： Human osteosarcoma cell line Saos-2 were c altured in vivo, Notchl signaling in Saos-2 cells was activated using ICN1 （Notch1 intracellular domain, tile active form of Notch1 signaling） transfection. Cell invasion ability was detected using a transwell system. The mRNA expression levels of RANKL,RANK, and OPG were analyzed using Real-time PCR. Results： Notch1 signaling was activated in Saos-2 ceils. Cell invasion number was 93. 7± 12. 8 and 41.4± 6. ; in Saos-2-ICN and Saos-2-G418, respectively. Relative mRNA expression levels of RANKL, OPG, and RANK in Saos-2- G418 were 0. 60±0.07, 1.2±0.14, and 0.3±0.04; in Saos-2-ICN cells, the corresponding levels were 1.8±0.3, 0.58±0.09, and 2.0±0.4 showing significant difference between the two groups （P〈0.05）. Oonclusions. Notchl activation increases cell invasion ability and mRNA expression level of RANKL and RANK; while down-regulates OPG expression level in Saos-2 cells. Results of this study suggest that RANKL-RANK-OPG could play a role in Notchl-induced osteosarcoma cell invasion and metastasis ability.
出处
《海南医学院学报》
CAS
2012年第9期1153-1156,共4页
Journal of Hainan Medical University
基金
国家自然科学基金(No.81000460)
海南省自然科学基金(No.309040)~~
