Chen Nafei, Meng Zhen, Song Jiaojie, Kong Lingfang, Zhang Yehua, Guo Suli, Zhang Xiaokun, Lu Xin, Jiang Licai, Chen Ran, Jiao Zongjiu, Zhao Liyun
Department of Hematology, Xingtai People's Hospital Xingtai 054000, Hebei Province, China.
Department of Hematology, Hudson International Peace Hospital, Heng Shui City People's Hospital Hengshui 053000, Hebei Province, China.
Am J Transl Res. 2021 Aug 15;13(8):9278-9284. eCollection 2021.
To validate the role of miR-497-5p in apoptosis in K562 cells by targeting Rho-associated kinase isoform 1 (ROCK1).
From January, 2017 to February, 2019, 57 patients with chronic myeloid leukemia (CML) treated in our hospital were included in patient group, and 50 healthy individuals were recruited as control group. miR-497-5p level in peripheral blood was quantitated using qRT-PCR. After transfecting with miR-497-5p overexpression vector and ROCK1 inhibitor, K562 cells were monitored in terms of proliferation (CCK8 assay), migration and invasion (Transwell), and apoptosis (flow cytometry). Binding loci between miR-497-5p and ROCK1 were predicted, and the targeting relationship was confirmed (dual-luciferase reporter (DLR) assay).
miR-497-5p was poorly expressed in CML ( < 0.05). Forced overexpression of miR-497-5p or inhibition of ROCK1 suppressed malignant processes (proliferation, proliferation, migration and invasion) in K562 cells and induced apoptosis ( < 0.05). DLR assay revealed a decreased luciferase activity after miR-497-5p binding to ROCK1 ( < 0.05).
miR-497-5p induces apoptosis in K562 cells by downregulating ROCK1.
通过靶向Rho相关激酶同工型1（ROCK1）来验证miR-497-5p在K562细胞凋亡中的作用。
2017年1月至2019年2月，将我院收治的57例慢性髓性白血病（CML）患者纳入患者组，招募50名健康个体作为对照组。采用qRT-PCR定量外周血中miR-497-5p水平。用miR-497-5p过表达载体和ROCK1抑制剂转染后，通过CCK8法检测K562细胞的增殖情况，通过Transwell检测其迁移和侵袭能力，通过流式细胞术检测其凋亡情况。预测miR-497-5p与ROCK1之间的结合位点，并通过双荧光素酶报告基因（DLR）试验证实靶向关系。
miR-497-5p在CML中表达较低（<0.05）。miR-497-5p的强制过表达或ROCK1的抑制可抑制K562细胞的恶性进程（增殖、迁移和侵袭）并诱导凋亡（<0.05）。DLR试验显示miR-497-5p与ROCK1结合后荧光素酶活性降低（<0.05）。
miR-497-5p通过下调ROCK1诱导K562细胞凋亡。
