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目的研究miR-19a对血肿瘤屏障通透性的影响。方法将miR-19a模拟物转染至人脑微血管内皮细胞hCMEC/D3,应用real-timePCR法检测miR-19a的表达。用过表达miR-19a的hCMEC/D3细胞和人U251胶质瘤细胞建立体外血肿瘤屏障模型,跨内皮电阻测量系统检测血肿瘤屏障跨内皮阻抗值的变化;Western blot和免疫荧光法检测体外血肿瘤屏障hCMEC/D3细胞中,紧密连接相关蛋白ZO-1和occludin的表达。结果经miR-19a模拟物转染后,hCMEC/D3细胞中miR-19a的表达水平显著升高;血肿瘤屏障跨内皮阻抗值显著下降;体外血肿瘤屏障hCMEC/D3细胞中,紧密连接相关蛋白ZO-1和occludin的表达水平显著降低,在细胞膜上呈不连续分布。结论 miR-19a过表达能显著增加血肿瘤屏障的通透性,其机制之一可能与降低紧密连接相关蛋白相关。
Objective To study the effect of miR-19a on the barrier permeability of blood tumor. Methods miR-19a mimics were transfected into hCMEC / D3 human brain microvascular endothelial cells and the expression of miR-19a was detected by real-time PCR. The in vitro hematological tumor barrier model was established by hCMEC / D3 cells overexpressing miR-19a and human U251 glioma cells. The trans-endothelial impedance was used to measure the trans-endothelial impedance of the tumor. Western blot and immunofluorescence were used to detect the extracorporeal blood Tumor barrier hCMEC / D3 cells, tight junction related protein ZO-1 and occludin expression. Results The expression of miR-19a in hCMEC / D3 cells was significantly increased after transfection with miR-19a mimics; the trans-endothelial impedance was significantly decreased in the hematological tumor barrier; in the hCMEC / D3 cells in vitro, the hCMEC / D3 cells were closely linked to related proteins The expression levels of ZO-1 and occludin were significantly decreased and discontinuous in the cell membrane. Conclusion Overexpression of miR-19a can significantly increase the permeability of blood-borne tumor barrier. One of the mechanisms may be related to the reduction of tight junction-related proteins.