摘要
目的构建白血病细胞黏附介导耐药(celladhesionmediateddrugresistance,CAM-DR)模型,从一新的角度研究白血病细胞耐药机制。方法采用人淋巴细胞白血病Jurkat细胞株与经60Co射线照射后的白血病骨髓基质细胞贴壁层共培养构建CAM-DR模型,应用扫描电镜从形态及结构对该模型进行鉴定;MTT法测定柔红霉素(daunorubicin,DNR)在Jurkat细胞中的IC50及运用流式细胞仪检测DNR在Jurkat细胞内的储积浓度及分布;AnnexinⅤ-FITC/PI双参数法流式细胞仪定量Jurkat细胞凋亡率。结果Jurkat细胞与骨髓基质细胞共培养24h,即观察到Jurkat细胞通过伪足黏附于骨髓基质细胞层,部分Jurkat细胞通过伪足龛于骨髓基质细胞融合所形成的“网眼”中,培养48h时可见部分Jurkat细胞移行到骨髓基质细胞层下,包裹于骨髓基质细胞层内,并可见Jurkat细胞呈巢状龛于骨髓基质细胞“网眼”中。该模型未影响DNR在Jurkat细胞中的蓄积及分布,但显著抑制了DNR诱导Jurkat细胞的凋亡及Jurkat细胞对DNR的反应性。结论成功构建了CAM-DR模型,从一新的角度为进一步研究白血病细胞耐药机制奠定了基础。
Objective To study the mechanism of drug resistance of leukemic cells in a new way by constructing cell adhesion mediated drug resistance (CAM-DR) model of leukemia. Methods Firstly, Jurkat cell strain of human lymphocytic leukemia were cocultured with the parietal layer of leukemic marrow matrix cells (BMSCs) radiated with ^60Co to construct the model of CAM-DR, then the model was assessed in morph and constructure by scanning electron microscope. The IC50 of daunorubicin (DNR) in Jurkat cells was examined by MTT and the concentration and distribution of DNR in Jurkat cells were detected by flow cytometry. The apoptosis ratio of Jurkat cells was quantitated by flow cytometry with double parameter methods with Annexin V -FITC/ PI. Results When the Jurkat cells were cocultured with BMSCs for 24 h, the Jurkat cells adhered to the matrix cell layer by parapodium and some of Jurkat cells had niched the mesh formed by the confluence of BMSCs ; at 48 h, some Jurkat cells had migrated to underlayer of BMSCs and were packaged, and Jurkat cells like nest niched the mesh of BMSCs. The accumulation and distribution of DNR in Jurkat cells were not affected in the model, but the apoptosis induced by DNR and the reaction of Jurkat cells to DNR were significantly inhibited. Conclusion The model of CAM-DR was successfully built, laying the foundation for further study on the mechanism of drug resistance of leukemia from a new viewpoint.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2006年第23期2336-2339,共4页
Journal of Third Military Medical University
基金
国家自然科学基金资助项目(39770335)~~
关键词
CAM—DR
模型
耐药
黏附
共培养
cell adhesion mediated drug resistance
model
drug resistance
adhesion
coculture