Department of Bone and Soft Tissue Tumor, Tianjin Medical University Cancer Hospital & Institute, Tianjin 300060, China.
BMC Med Genomics. 2013 Dec 15;6:56. doi: 10.1186/1755-8794-6-56.
To investigate associations between WW domain-containing oxidoreductase (WWOX), runt-related transcription factor 2 (RUNX2) and vascular endothelial growth factor alpha (VEGFA) in human osteosarcoma (OS).
Copy number aberrations of WWOX, RUNX2and VEGFA genes were detected by microarray comparative genomic hybridization (aCGH) in 10 fresh OS tissue samples. VEGFA gene alterations were also investigated and validated by fluorescence in situ hybridization (FISH) in 54 formalin-fixed and paraffin-embedded (FFPE) OS samples. Protein expression of WWOX, RUNX2 and VEGFA were examined in 54 FFPE OS samples by immunohistochemistry (IHC).
Analysis of previously published OS aCGH data (GSE9654) and aCGH data from this study (GSE19180) identified significant deletion of WWOX in 30% (6/20) of OS samples, whilst significant increase in both RUNX2 and VEGFA gene copy numbers were detected in 55% (11/20) and 60% (12/20) of OS samples, respectively. FISH demonstrated increased VEGFA gene copy number in 65.9% (31/47) of evaluable samples, in either focal or large fragment forms. Compared with positive expression of WWOX in 38.9% of the OS samples, positive expression of RUNX2 and VEGFA protein was found in 48.1 and 75.9% of samples. Although there was no significant association between gene copy number aberration and protein expression for WWOX and RUNX2, significant positive correlation between increased VEGFA gene copy number and VEGFA protein expression was observed. Although there was no significant reverse association between WWOX and RUNX2 expression, a significantly positive relationship was observed between RUNX2 and VEGFA protein expression.
Our data show increased RUNX2 and VEGFA gene copy numbers and elevation of their respective proteins in human OS. Positive correlation of RUNX2 and VEGFA suggests that both increased VEGFA gene copy number and RUNX2 overexpression facilitate increased expression of VEGFA.
研究 WW 结构域包含的氧化还原酶（WWOX）、 runt 相关转录因子 2（RUNX2）和血管内皮生长因子 alpha（VEGFA）在人骨肉瘤（OS）中的关联。
通过微阵列比较基因组杂交（aCGH）在 10 例新鲜 OS 组织样本中检测 WWOX、RUNX2 和 VEGFA 基因的拷贝数异常。在 54 例福尔马林固定和石蜡包埋（FFPE）OS 样本中通过荧光原位杂交（FISH）也研究和验证了 VEGFA 基因的改变。通过免疫组织化学（IHC）检测 54 例 FFPE OS 样本中 WWOX、RUNX2 和 VEGFA 的蛋白表达。
分析先前发表的 OS aCGH 数据（GSE9654）和本研究的 aCGH 数据（GSE19180），发现 30%（6/20）的 OS 样本中存在 WWOX 的显著缺失，而 RUNX2 和 VEGFA 基因拷贝数的显著增加分别在 55%（11/20）和 60%（12/20）的 OS 样本中检测到。FISH 显示在 65.9%（31/47）可评估样本中存在 VEGFA 基因拷贝数增加，表现为局灶性或大片段形式。与 OS 样本中 WWOX 阳性表达的 38.9%相比，RUNX2 和 VEGFA 蛋白的阳性表达分别在 48.1%和 75.9%的样本中发现。尽管 WWOX 和 RUNX2 的基因拷贝数异常与蛋白表达之间没有显著关联，但 VEGFA 基因拷贝数的增加与 VEGFA 蛋白表达之间存在显著的正相关。尽管 WWOX 和 RUNX2 表达之间没有显著的反向关联，但观察到 RUNX2 和 VEGFA 蛋白表达之间存在显著的正相关关系。
我们的数据显示，人骨肉瘤中 RUNX2 和 VEGFA 基因的拷贝数增加，相应蛋白水平升高。RUNX2 和 VEGFA 的正相关表明，VEGFA 基因拷贝数的增加和 RUNX2 的过表达都促进了 VEGFA 的表达。
