丝裂原活化蛋白激酶信号通路相关基因在人骨肉瘤中的表达

Gene profiling of MAPK pathway in human osteosarcoma

目的探讨丝裂原活化蛋白激酶（MAPK）信号通路相关基因在人骨肉瘤发病过程中的作用。方法应用Affymetrix的Human Genome U133A芯片检测骨肉瘤细胞与成骨细胞间MAPK信号通路相关基因表达谱的变化，然后利用MATLAB软件对芯片检测到的差异表达明显的基因进行KEGG通路分析。采用免疫组织化学技术检测48例骨肉瘤和24例成骨性良性肿瘤组织中ERK1／2、JNK和p38蛋白的表达。结果以表达差异≥2倍为限，骨肉瘤细胞株MG-63、Saos-2和U-2OS中，共筛选出18个与成骨细胞株hFOB1．19中存在差异表达的MAPK信号通路相关基因，其中10个为上调基因，8个为下调基因。18个差异表达的基因在KEGG的MAPK信号通路上均为重要的节点基因。免疫组织化学染色结果显示，ERK1／2、JNK和p38蛋白在骨肉瘤组织中的阳性表达率分别为83．3％（40／48）、72．9％（35／48）和85．4％（41／48），在成骨性良性肿瘤组织中的阳性表达率分别为12．5％（3／24）、8．3％（2／24）和16．7％（4／24），差异均有统计学意义（均P〈0．01）。结论MAPK信号通路在骨肉瘤的发生中发挥着重要作用，其中ERK1／2、JNK和p38三条通路通过相互协调形成复杂的调节网络，参与调节骨肉瘤细胞的增殖、分化和凋亡，以及骨肉瘤细胞的侵袭和转移。

Objective To explore the functional effetcs of MAPK pathway in the pathogenesis of human osteosarcoma. Methods Gene microarray （Human Genome U133A, Affymetrix ） was used to screen the differential expression of genes involved in MAPK pathway between osteosarcoma cell lines and 3 osteoblastic cell lines. KEGG metabolic pathway analysis was performed among significantly increased or decreased genes using the MATLAB software. Immunohistochemical technique was used to detect the expressions of ERK1/2, JNK and p38 proteins among 48 osteosarcoma and benign 24 osteoblastic tumor samples. Results Using an entrance limit of ≥2.0, 18 differentially expressed MAPK pathway-related genes were selected （10 up-regulated, 8 down-regulated） to mapped to the MAPK pathway of KEGG which are all important node genes. The positive rates of ERK1/2, JNK and p38 proteins were 83.3% （40/48） , 72.9% （35/48） and 85.4% （41/48） in osteosarcomas,and 12.5% （3/24）,8.3% （2/24） and 16.7% （4/24） in the control group, respectively. The positive rates and expression intensities were statistically different between the 2 groups （ P 〈 0.01 ）. Conclusion MAPK pathway plays an important role in the pathogenesis of osteosarcoma. ERK, JNK and p38 form an intercoordinating network and regulate the cell proliferation, differentiation, apoptosis, invasion and migration in osteosarcoma.