Interaction of cytokines and cell adhesion molecules during osteoclastogenesis

大多数类型的细胞需要与细胞外基质接触，而细胞外基质主要由整合素介导。整合素的信号转导途径与细胞周期蛋白的信号转导途径有许多共同之处。破骨细胞前体细胞表达丰富的整合素av-β-5，需要多种细胞因子包括M-CSF5才能成熟和存活。因此，我们探索了整合素和细胞因子信号通路对破骨细胞前体的相互作用。从C3H小鼠后肢骨分离破骨前体细胞，在PFA培养皿中加入M-CSF培养3天。收集破骨细胞前体细胞，在含有(+)或不含(-)M-CSF的塑料(A)或PFD(NA)培养皿中培养。各组细胞裂解产物经SDS-PAGE分级、印迹、抗磷酸化酪氨酸鉴定，50 kDa条带受细胞附着调控，NA较A减少，NA-与NA+间M-CSF作用不明显，A+50 kDa条带强度是A-的1.3倍，50 kDa蛋白是信号转导分子的候选者。细胞黏附是M-CSF发挥作用所必需的，因为M-CSF上调了A。

Most type of cells need to contact with extracellular matrix, which are mainly mediated with integrins. The signal transduction pathway through integrin shares many parts with that of cyokines. The expression of integrin avβ5 is abundant on osteoclast precursors, which need many kinds of cyokines, including M-CSF, for their maturation and survival. Therefore we explored interaction of integrin and cytokine signal pathway on osteoclast precursors.Osteoclast precursors were isolated from C3H mouse hindlimb bones and cultured in PFA dishes for 3 days with M-CSF. Osteoclast precursors were collected and cultured on plastic(A) or PFD(NA) dishes with(+) or without(-) M-CSF. Cell lysate from each group were fractionated on SDS-PAGE, blotted onto membrane and proved by anti-phosphorlated tyrosine.The 50kDa band were regulated by cell attachment and decreased in NA compared with A. The effect of M-CSF was not clear between NA- and NA+, the intensity of 50kDa band in A+ showed 1.3fold increase of that in A-.The 50kDa protein is the candidate of signal transduction molecules. Cell adhesion is necessary for the effect of M-CSF since the intensity of 50kDa band was up-regulated by M-CSF in A.