Objective: To detect the differential expressions of miR-451, ABCB1 and ABCC2 in drug-sensitive leukemia cell line K562 and drug-resistant cell line K562/A02, and explore the regulatory relationship between miR-451 and the expressions of ABCB1 and ABCC2 , and the mechanism of miR-451 involved in drug resistance in leukemia.
Methods: CCK-8 assay was used to detect the drug resistance of K562/A02 and K562 cells. Quantitative Real-time PCR (qRT-PCR) was used to verify the differential expressions of miR-451 in K562 and K562/A02 cells. MiR-451 mimic and negative control (miR-NC), miR-451 inhibitor and negative control (miR-inNC) were transfected into K562 and K562/A02 cells respectively, then qRT-PCR and Western blot were used to detect the expression levels of mRNA and protein of ABCB1 and ABCC2 in K562 and K562/A02 cells and the transfected groups.
Results: The drug resistance of K562/A02 cells to adriamycin was 177 times higher than that of its parent cell line K562. Compared with K562 cells, the expression of miR-451 in K562/A02 cells was significantly higher (P <0.001), and the mRNA and protein expression levels of ABCB1 and ABCC2 in K562/A02 cells were significantly higher than those in K562 cells (P <0.001). After transfected with miR-451 inhibitor, the expression of miR-451 was significantly down-regulated in K562/A02 cells (P <0.001), the sensitivity to chemotherapy drugs was significantly enhanced (P <0.05), and the mRNA and protein expressions of ABCB1 and ABCC2 were significantly decreased (P <0.01). After transfected with miR-451 mimic, the expression of miR-451 was significantly upregulated in K562 cells (P <0.001), and the mRNA and protein expressions of ABCB1 and ABCC2 were significantly increased (P <0.01).
Conclusion: There are significant differences in the expressions of miR-451, ABCB1 and ABCC2 between the drug-sensitive leukemia cell line K562 and drug-resistant cell line K562/A02, which suggests that miR-451 may affect the drug resistance of leukemia cells by regulating the expression of ABCB1 and ABCC2.
题目: miR-451在白血病细胞株K562/A02多药耐药中的作用及相关机制.
目的: 检测miR-451、ABCB1 、ABCC2 在白血病药物敏感细胞株K562及其耐药株K562/A02中的差异表达，探讨miR-451与ABCB1 、ABCC2 表达的调控关系及miR-451参与白血病耐药的相关机制。.
方法: CCK-8法检测K562/A02及K562细胞的耐药性，实时荧光定量PCR（qRT-PCR）验证miR-451在K562及K562/A02细胞中的差异表达，将miR-451模拟物（mimic）及其阴性对照（miR-NC）、miR-451抑制剂（inhibitor）及其阴性对照（miR-inNC）分别转染K562及K562/A02细胞，应用qRT-PCR、Western blot检测ABCB1 、ABCC2 在K562、K562/A02细胞及转染后各组细胞中的mRNA、蛋白表达水平。.
结果: K562/A02细胞对阿霉素的耐药是其亲本细胞系K562的177倍。与K562细胞相比，K562/A02细胞中miR-451明显高表达（P <0.001），ABCB1 、ABCC2 在K562/A02中的mRNA及蛋白表达水平均显著高于K562细胞（P <0.001）。K562/A02细胞转染miR-451 inhibitor后，miR-451表达显著下调（P <0.001），对化疗药物的敏感性显著增强（P <0.05），ABCB1 、ABCC2 mRNA及蛋白表达水平显著降低（P <0.01）。K562细胞转染miR-451 mimic后，miR-451表达显著上调（P <0.001），ABCB1 、ABCC2 mRNA及蛋白表达水平显著增高（P <0.01）。.
结论: miR-451、ABCB1 及ABCC2 在白血病敏感细胞株K562及其耐药株K562/A02中的表达存在显著差异，miR-451可能通过调控ABCB1 、ABCC2 的表达对白血病细胞的耐药性产生影响。.
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