Hu Yingwei, Yu Shifeng
Department of Oral Pathology, School of Stomatology, Peking University, Beijing 100081, China.
Zhonghua Bing Li Xue Za Zhi. 2002 Apr;31(2):128-31.
To investigate the gene expression of osteoprotegerin (OPG) and osteoclast differentiation factor (ODF/TRANCE/RANKL), two new members of the TNF-receptor superfamily, in giant cell tumor (GCT); to discuss the molecular mechanism of extensive bone resorption caused by GCT.
Using TRIzol reagent to prepare total RNA from GCT sample and normal bone tissue. By a first-strand complementary DNA (cDNA) synthesis kit, cDNA was synthesized from 2.0 micro g RNA according to the manufacturer's instructions. cDNA was then amplified by PCR. Amplification products were resolved by electrophoresis on a 1.5% agarose gel and stained with EB. The relative quantity of the PCR products were determined and the mRNA levels of OPG, ODF, M-CSF (cofactor of ODF), and RANK (receptor of ODF) were compared with that of the normal bone.
GCT contained highly expressed mRNA of ODF, OPG, M-CSF and RANK. There was mRNA expression of OPG, M-CSF and RANK and less expression of ODF in normal bone. The ODF mRNA and RANK mRNA in GCT were more abundant than that in normal bone. In GCT, the ratio of ODF mRNA exceeded OPG expression. But in normal bone, the OPG mRNA exceeded ODF expression.
The results suggest that GCT contains all signals including OPG, ODF, M-CSF and RANK that are essential for inducing osteoclastogenesis and promoting bone resorption.
研究肿瘤坏死因子受体超家族的两个新成员骨保护素(OPG)和破骨细胞分化因子(ODF/TRANCE/RANKL)在骨巨细胞瘤(GCT)中的基因表达;探讨GCT引起广泛骨吸收的分子机制。
用TRIzol试剂从GCT样本和正常骨组织中提取总RNA。使用第一链互补DNA(cDNA)合成试剂盒,按照制造商说明从2.0μg RNA合成cDNA。然后通过PCR扩增cDNA。扩增产物在1.5%琼脂糖凝胶上进行电泳分离,并用EB染色。测定PCR产物的相对量,并将OPG、ODF、M-CSF(ODF的辅助因子)和RANK(ODF的受体)的mRNA水平与正常骨的进行比较。
GCT中ODF、OPG、M-CSF和RANK的mRNA高表达。正常骨中有OPG、M-CSF和RANK的mRNA表达,ODF表达较少。GCT中的ODF mRNA和RANK mRNA比正常骨中更丰富。在GCT中,ODF mRNA的比例超过OPG表达。但在正常骨中,OPG mRNA超过ODF表达。
结果表明,GCT包含诱导破骨细胞生成和促进骨吸收所必需的所有信号,包括OPG、ODF、M-CSF和RANK。