Yang Ru, Ma Dong, Wu Yanwei, Zhang Yingzi, Zhang Lina
Henan Key Laboratory of Neurorestoratology, First Affiliated Hospital of Xinxiang Medical University, Weihui, Henan Province, People's Republic of China.
Hematology Laboratory, First Affiliated Hospital of Xinxiang Medical University, Weihui, Henan Province, People's Republic of China.
Onco Targets Ther. 2020 Dec 17;13:12943-12954. doi: 10.2147/OTT.S258684. eCollection 2020.
At present, there is a lack of precise knowledge on acute myeloid leukemia (AML) at the molecular level, and understanding its occurrence at the genetic level is conducive to the development of targeted therapies. Therefore, in this study the relationship between the lncRNA -miR183-5p-FOXO1 axis and AML was explored.
Expression of lncRNA and miR183-5p was quantified by quantitative real-time PCR, and the level of FOXO1 and other proteins was measured by Western blot. Expression vectors of lncRNA , miR183-5p, and FOXO1 were constructed to assess effects of the three on cell proliferation and apoptosis. MTT reduction assays were employed for cell proliferation, flow cytometry for cell cycle and apoptosis, and dual luciferase-reporter assays for the targeting relationship between lncRNA and miR183-5p and miR183-5p and FOXO1.
lncRNA was highly expressed in peripheral blood/leukemia cell lines of patients with AML compared with normal human peripheral blood/peripheral blood mononuclear cells. miR183-5p was the target of lncRNA and the target gene of miR183-5p rather than lncRNA SNHG16. Absence of lncRNA led to upregulation of miR183-5p, promotion of apoptosis, and inhibition of proliferation. Suppression of miR183-5p accelerated cell proliferation and hindered apoptosis. miR183-5p negatively regulated FOXO1, and FOXO1 promoted proliferation and inhibited apoptosis. Inhibition of miR183-5p counteracted the changes caused by lncRNA absence.
lncRNA regulates the progress of AML via the miR183-5p-FOXO1 axis.
目前，在分子水平上对急性髓系白血病（AML）缺乏精确的认识，而了解其在基因水平上的发生有助于靶向治疗的发展。因此，本研究探讨了lncRNA -miR183-5p-FOXO1轴与AML之间的关系。
通过定量实时PCR定量lncRNA和miR183-5p的表达，通过蛋白质免疫印迹法检测FOXO1等蛋白的水平。构建lncRNA、miR183-5p和FOXO1的表达载体，以评估三者对细胞增殖和凋亡的影响。采用MTT比色法检测细胞增殖，流式细胞术检测细胞周期和凋亡，双荧光素酶报告基因检测法检测lncRNA与miR183-5p以及miR183-5p与FOXO1之间的靶向关系。
与正常人外周血/外周血单个核细胞相比，lncRNA在AML患者外周血/白血病细胞系中高表达。miR183-5p是lncRNA的靶标，且是miR183-5p而非lncRNA SNHG16的靶基因。lncRNA缺失导致miR183-5p上调、促进凋亡并抑制增殖。抑制miR183-5p可加速细胞增殖并阻碍凋亡。miR183-5p负向调节FOXO1，而FOXO1促进增殖并抑制凋亡。抑制miR183-5p可抵消lncRNA缺失所引起的变化。
lncRNA通过miR183-5p-FOXO1轴调节AML的进展。
