Feng Yan-Li, Su Bao-Xiong, Ge Fan-Mei, Dai Chong-Wen
Department of Hematology, Yan'an University Affiliated Hospital, Yan'an 716000, Shaanxi Province, China.
Department of Hematology, The Second Xiangya Hospital of Central South University, Changsha 410011, Hunan Province, China.E-mail:
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2023 Jun;31(3):685-692. doi: 10.19746/j.cnki.issn.1009-2137.2023.03.011.
To detect the differential expressions of miR-451, and 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 and , and the mechanism of miR-451 involved in drug resistance in leukemia.
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 and in K562 and K562/A02 cells and the transfected groups.
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 ( <0.001), and the mRNA and protein expression levels of and in K562/A02 cells were significantly higher than those in K562 cells ( <0.001). After transfected with miR-451 inhibitor, the expression of miR-451 was significantly down-regulated in K562/A02 cells ( <0.001), the sensitivity to chemotherapy drugs was significantly enhanced ( <0.05), and the mRNA and protein expressions of and were significantly decreased ( <0.01). After transfected with miR-451 mimic, the expression of miR-451 was significantly upregulated in K562 cells ( <0.001), and the mRNA and protein expressions of and were significantly increased ( <0.01).
There are significant differences in the expressions of miR-451, and 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 and .
检测miR-451在白血病药敏细胞株K562和耐药细胞株K562/A02中的差异表达，探讨miR-451与[未提及的两个基因，原文中此处缺失]表达之间的调控关系以及miR-451参与白血病耐药的机制。
采用CCK-8法检测K562/A02和K562细胞的耐药性。运用定量实时荧光定量PCR（qRT-PCR）验证miR-451在K562和K562/A02细胞中的差异表达。将miR-451模拟物和阴性对照（miR-NC）、miR-451抑制剂和阴性对照（miR-inNC）分别转染至K562和K562/A02细胞中，然后采用qRT-PCR和蛋白质免疫印迹法检测K562和K562/A02细胞及转染组中[未提及的两个基因，原文中此处缺失]的mRNA和蛋白表达水平。
K562/A02细胞对阿霉素的耐药性是其亲本细胞株K562的177倍。与K562细胞相比，K562/A02细胞中miR-451的表达显著升高（P<0.001），且K562/A02细胞中[未提及的两个基因，原文中此处缺失]的mRNA和蛋白表达水平显著高于K562细胞（P<0.001）。转染miR-451抑制剂后，K562/A02细胞中miR-451的表达显著下调（P<0.001），对化疗药物的敏感性显著增强（P<0.05），[未提及的两个基因，原文中此处缺失]的mRNA和蛋白表达显著降低（P<0.01）。转染miR-451模拟物后，K562细胞中miR-451的表达显著上调（P<0.001），[未提及的两个基因，原文中此处缺失]的mRNA和蛋白表达显著增加（P<0.01）。
白血病药敏细胞株K562和耐药细胞株K562/A02中miR-451、[未提及的两个基因，原文中此处缺失]的表达存在显著差异，提示miR-451可能通过调控[未提及的两个基因，原文中此处缺失]的表达影响白血病细胞的耐药性。
