Jin Jing, Yao Jia, Yue Fang, Jin Zhaoying, Li Dan, Wang Shan
Department of Pharmacy, Jining No. 1 People's Hospital, Jining, Shandong 272011, P.R. China.
Department of Pharmacy, Qilu Medical University, Zibo, Shandong 255213, P.R. China.
Exp Ther Med. 2018 Sep;16(3):1693-1700. doi: 10.3892/etm.2018.6404. Epub 2018 Jul 4.
The aim of the present study was to determine the expression of adenosine triphosphate binding cassette subfamily B member 1 (ABCB1) gene and its protein P-glycoprotein (PGP) in bone marrow mononuclear cells from chronic myeloid leukemia (CML) patients with imatinib mesylate (IM) resistance, or IM-resistant CML K562 cells. In addition, the molecular mechanism of action of microRNA (miR)-214 on ABCB1 in IM resistance was investigated. A total of 26 CML patients with IM resistance were included in the present study. In addition, 31 CML patients who did not have IM resistance were included as the control group. Bone marrow was collected from all subjects. The K562R cell line, which is a K562 cell line with IM resistance, was used for cellular studies. Reverse transcription-quantitative polymerase chain reaction was used to determine the expression of ABCB1 mRNA and miR-214 in cells. Western blotting was employed to determine the expression of PGP. Dual luciferase reporter assay was carried out to identify interactions between ABCB1 mRNA and miR-214. MTT assay was used to determine the survival rate of cells. ABCB1 mRNA and PGP expression was upregulated in bone marrow mononuclear cells from CML patients with IM resistance. K562R cells had higher ABCB1 and PGP expression than K562 cells, potentially due to their different sensitivity to IM. Expression miR-214 was decreased in bone marrow mononuclear cells from patients with IM resistance and K562R cells. Notably, miR-214 was able to bind with the 3'-untranslated region, seed region of ABCB1 mRNA to regulate its expression. In addition, elevated expression of miR-214 restored IM sensitivity to K562R cells potentially by affecting ABCB1 expression. The present study demonstrated that upregulated expression of ABCB1 mRNA and PGP in bone marrow mononuclear cells from CML patients with IM resistance may be associated with the downregulation of miR-214. In addition, miR-214 may participate in the IM resistance of CML patients by regulating ABCB1 expression.
本研究的目的是确定慢性髓性白血病（CML）甲磺酸伊马替尼（IM）耐药患者或IM耐药的CML K562细胞的骨髓单个核细胞中三磷酸腺苷结合盒亚家族B成员1（ABCB1）基因及其蛋白P-糖蛋白（PGP）的表达。此外，还研究了微小RNA（miR）-214在IM耐药中对ABCB1的分子作用机制。本研究共纳入26例IM耐药的CML患者。此外，纳入31例无IM耐药的CML患者作为对照组。采集所有受试者的骨髓。K562R细胞系是一种对IM耐药的K562细胞系，用于细胞研究。采用逆转录定量聚合酶链反应测定细胞中ABCB1 mRNA和miR-214的表达。采用蛋白质免疫印迹法测定PGP的表达。进行双荧光素酶报告基因检测以鉴定ABCB1 mRNA与miR-214之间的相互作用。采用MTT法测定细胞存活率。IM耐药的CML患者骨髓单个核细胞中ABCB1 mRNA和PGP表达上调。K562R细胞比K562细胞具有更高的ABCB1和PGP表达，这可能是由于它们对IM的敏感性不同。IM耐药患者的骨髓单个核细胞和K562R细胞中miR-214表达降低。值得注意的是，miR-214能够与ABCB1 mRNA的种子区域3'-非翻译区结合以调节其表达。此外，miR-214表达升高可能通过影响ABCB1表达恢复K562R细胞对IM的敏感性。本研究表明，IM耐药的CML患者骨髓单个核细胞中ABCB1 mRNA和PGP表达上调可能与miR-214下调有关。此外，miR-214可能通过调节ABCB1表达参与CML患者的IM耐药。
