CONNEXIN43 MEDIATES DIRECT INTERCELLULAR COMMUNICATION IN HUMAN OSTEOBLASTIC CELL NETWORKS
CONNEXIN43 MEDIATES DIRECT INTERCELLULAR COMMUNICATION IN HUMAN OSTEOBLASTIC CELL NETWORKS
复制标题
DOI：
10.1172/jci116406
复制
发表时间：
1993-05-01
影响因子：
15.9
通讯作者：
STEINBERG, TH
中科院分区：
文献类型：
--
作者：
CIVITELLI, R;BEYER, EC;STEINBERG, TH
我们在人骨髓基质细胞（BMC）和骨小梁成骨细胞（HOB）以及人成骨肉瘤细胞系SaOS-2的单层培养细胞中检测了细胞偶联和间隙连接蛋白的表达。由于微量注射路西法黄导致染料转移到邻近细胞，HOB和BMC细胞都是功能偶联的，平均每次注射分别有3.4+/-2.8 （n = 131）和8.1+/-9.3 （n = 51）个偶联细胞。相反，在SaOS-2单层细胞中，Lucifer黄的扩散很少（每次注射1.4个+/-1.8个偶联细胞，n = 100）。辛醇（3.8 mM）抑制了染料的扩散，辛醇是一种间隙连接通讯抑制剂。所有成骨细胞均表达连接蛋白43和连接蛋白45的mRNA，但不表达连接蛋白26、32、37、40和46。尽管所有成骨细胞表达相似数量的连接蛋白43 mRNA，但通过免疫荧光和免疫沉淀评估，低偶联的SaOS-2细胞产生的Cx43蛋白明显少于HOB或BMC。相反，SaOS-2细胞比HOB或BMC细胞表达更多的Cx45 mRNA。因此，正常和转化的人成骨细胞中的细胞间偶联与Cx43的表达水平相关，而Cx43似乎介导了这些细胞中的细胞间通讯。缝隙连接通信可以作为成骨细胞同步工作并将局部产生的信号传播到整个骨组织的一种手段。
We have examined cell coupling and expression of gap junction proteins in monolayer cultures of cells derived from human bone marrow stromal cells (BMC) and trabecular bone osteoblasts (HOB), and in the human osteogenic sarcoma cell line, SaOS-2. Both HOB and BMC cells were functionally coupled, since microinjection of Lucifer yellow resulted in dye transfer to neighboring cells, with averages of 3.4+/-2.8 (n = 131) and 8.1+/-9.3 (n = 51 ) coupled cells per injection, respectively. In contrast, little diffusion of Lucifer yellow was observed in SaOS-2 monolayers (1.4+/-1.8 coupled cells per injection, n = 100). Dye diffusion was inhibited by octanol (3.8 mM), an inhibitor of gap junctional communication. All of the osteoblastic cells expressed mRNA for connexin43 and connexin45, but not for connexins 26, 32, 37, 40, or 46. Whereas all of the osteoblastic cells expressed similar quantities of mRNA for connexin43, the poorly coupled SaOS-2 cells produced significantly less Cx43 protein than either HOB or BMC, as assessed by immunofluorescence and immunoprecipitation. Conversely, more Cx45 mRNA was expressed by SaOS-2 cells than by HOB or BMC. Thus, intercellular coupling in normal and transformed human osteoblastic cells correlates with the level of expression of Cx43, which appears to mediate intercellular communication in these cells. Gap junctional communication may serve as a means by which osteoblasts can work in synchrony and propagate locally generated signals throughout the skeletal tissue.