Interaction between Osteopontin on Madin Darby Canine Kidney Cell Membrane and Calcium Oxalate Crystal
Interaction between Osteopontin on Madin Darby Canine Kidney Cell Membrane and Calcium Oxalate Crystal
复制标题
Madin Darby犬肾细胞膜上的骨桥蛋白与草酸钙晶体的相互作用
作者：
T. Yamate;K. Kohri;T. Umekawa;E. Konya;Y. Ishikawa;M. Iguchi;T. Kurita
我们最近报道了骨桥蛋白（OPN）的添加导致Madin Darby犬肾（MDCK）细胞表面草酸钙（CaOx）晶体沉积的增加。为了确定这种增加的沉积在多大程度上是由OPN引起的，在确定CaOx晶体结合水平之前，我们研究了由OPN在细胞表面表达产生的CaOx晶体沉积在多大程度上被4种不同的方法抑制。将MDCK细胞（2 × 106细胞/孔）培养至融合状态，加入人OPN多克隆抗体、凝血酶、环arg - gy - asp （RGD）肽和tunicamycin四种物质中的一种，抑制OPN与细胞表面的结合。细胞培养24 h后，采用荧光抗体技术结合OPN多克隆抗体检测细胞表面OPN的表达是否受到抑制，并采用同位素45Ca测定CaOx晶体沉积程度。抗体组CaOx晶体沉积程度抑制80%以上，凝血素组抑制50-80%，环rgd组抑制60-80%，tunicamycin组抑制50-60%。这些结果表明，细胞外基质中的OPN是MDCK细胞表面CaOx晶体沉积的主要原因。
We recently reported that the addition of the protein osteopontin (OPN) resulted in an increase in the deposition of calcium oxalate (CaOx) crystals on the surface of Madin Darby canine kidney (MDCK) cells. To determine the degree to which this increased deposition is caused by OPN, we investigated the extent to which the CaOx crystal deposition produced by the expression of OPN at the cell surface was suppressed by 4 different methods prior to the determination of the level of CaOx crystal binding. MDCK cells (2 × 106 cells/well) were cultured to a confluent state, and the binding of OPN to the cellular surface was then inhibited by adding one of the following 4 substances: human OPN polyclonal antibody, thrombin, cyclic Arg-Gly-Asp (RGD) peptides and tunicamycin. The cells were cultured for 24 h. We then used a fluorescent antibody technique with an OPN polyclonal antibody to determined whether the expression of OPN at the cell surface was inhibited, and we measured the degree of CaOx crystal deposition using the isotope 45Ca. The degree of CaOx crystal deposition was inhibited by 80% or more in the antibody-treated group, by 50–80% in the thrombin-treated group, by 60–80% in the cyclic RGD-treated group, and by 50–60% in the tunicamycin-treated group. These results suggest that OPN in the extracellular matrix is the main cause of CaOx crystal deposition on the surface of MDCK cells.
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影响因子：
--
作者：
LIESKE, JC;WALSHREITZ, MM;TOBACK, FG
通讯作者：
TOBACK, FG
DOI：
10.1073/pnas.91.15.6987
发表时间：
1994-07-19
影响因子：
11.1
作者：
LIESKE, JC;SWIFT, H;TOBACK, FG
通讯作者：
TOBACK, FG
DOI：
10.1016/0167-4838(89)90092-7
发表时间：
1989-06-13
期刊：
BIOCHIMICA ET BIOPHYSICA ACTA
影响因子：
--
作者：
SENGER, DR;PERRUZZI, CA;TENEN, DG
通讯作者：
TENEN, DG
影响因子：
3.2
作者：
MARK, MP;PRINCE, CW;BUTLER, WT
通讯作者：
BUTLER, WT