间隙连接蛋白Cx43磷酸化状态对白血病骨髓基质细胞GJIC功能的影响及作用机制研究

项目名称： 间隙连接蛋白Cx43磷酸化状态对白血病骨髓基质细胞GJIC功能的影响及作用机制研究

项目编号： No.81000194

项目类型： 青年科学基金项目

立项/批准年度： 2011

项目学科： 金属学与金属工艺

项目作者： 刘耀

作者单位： 中国人民解放军第三军医大学

项目金额： 10万元

中文摘要： 申请者在既往研究中发现白血病骨髓基质细胞GJIC功能较正常骨髓明显降低，但具体机制不明。连接蛋白翻译后的磷酸化调节是影响GJIC形成的重要因素，连接蛋白的磷酸化程度决定着GJIC的通透性及功能。因此推测白血病骨髓基质GJIC功能缺陷可能与基质细胞上Cx43的磷酸化状态异常有关。故体外培养正常和白血病骨髓基质细胞，观察二者Cx43磷酸化水平的差异，并干预急性白血病骨髓基质细胞异常的磷酸化状态，观察改变磷酸化状态后Cx43形成GJIC能力的变化；初步探讨通过干预Cx43磷酸化状态改善GJIC功能对白血病骨髓基质细胞生物学特性及造血功能的影响。研究发现：（1）急性白血病骨髓基质细胞Cx43的磷酸化水平高于正常骨髓基质细胞；在培养体系中加入磷酸酶抑制剂冈田酸后急性白血病骨髓基质细胞Cx43的磷酸化水平下降，去磷酸化水平增加；（2）阻断急性白血病骨髓基质细胞Cx43的过磷酸化，可以增强其形成GJIC的水平；（3）改变磷酸化状态对白血病骨髓基质细胞生物学特性和造血能力产生一定的影响。该研究初步阐明白血病造血微环境GJIC功能缺陷的机制，为改造白血病造血微环境提供新的思路和实验依据。

中文关键词： 间隙连接蛋白43；磷酸化；间隙连接细胞间通讯；骨髓基质细胞；白血病

英文摘要： The applicant find that compared with normal bone marrow the GJIC function of leukemic bone marrow stromal cells is significantly reduced in the previous study, but the specific mechanism is still unknown. Phosphorylation regulation of connexon after translation is an important factor in affecting GJIC, and the phosphorylation degree of connexon determines the permeability and function of GJIC. For this reason, we speculate that the GJIC function defects of leukemic bone marrow stromal may be related to phosphorylation abnormality of Cx43 in the stromal cells. Therefore, in vitro culture of normal and leukemic bone marrow stromal cells, observing the difference of the two kinds of Cx43 phosphorylation levels, intervening in the phosphorylation abnormality in acute leukemic bone marrow stromal cells, and observing after phosphorylation the ability of diversify in Cx43 formation of GJIC, we approach the intervening Cx43 phosphorylation to improve GJIC function of leukemic bone marrow stromal cells bionomics and influence of blood-producing function. The study discover that, firstly the phosphorylation level of acute leukemia bone marrow stromal cells Cx43 is higher than the normal bone marrow stromal cells, and after adding phosphatase enzyme inhibitors okadaic acid into cultivation system the phosphorylation level of acute leukemia bone marrow stromal cells Cx43 can be lower and dephosphorylation level can be increased,secondly blocking the phosphorylation process of acute leukemia bone marrow stromal cells Cx43 can enhance formation level of GJIC, thirdly changing the phosphorylation influence to leukemic bone marrow stromal cells bionomics and hematogenesis. This study clarity initially the mechanism of leukemia hematopoietic microenviroment GJIC functional defect, and provid new thread and test basis for reforming the leukemia hematopoietic microenviroment.

英文关键词： Cx43; phosphorytion; GJIC; bone marrow stromal cell; leukemia