Zhong Ling, Xu Zhuyu, Jin Xin, He Yuan, Zhang Jianbo, Jiang Tao, Chen Jiao
Department of Clinical Laboratory, Sichuan Academy of Medical Science & Sichuan Provincial People's Hospital, School of Medicine, University of Electronic Science and Technology of China, Chengdu, Sichuan 610072, P.R. China.
Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, Sichuan 610041, P.R. China.
Oncol Lett. 2020 Dec;20(6):339. doi: 10.3892/ol.2020.12202. Epub 2020 Oct 8.
The IL-6R/JAK2/STAT3 pathway mediated by interleukin-6 (IL-6) plays an important role in the occurrence and development of multiple myeloma (MM), which is associated with decreased microRNA-451a. However, the biological function of microRNA-451a in MM remains unclear. The bone marrow (BM) of patients with MM was sampled, and the plasma cells were enriched. BM miR-451a, IL-6 and IL-6R levels and Ki-67 expression intensity were evaluated using reverse transcription-quantitative PCR, ELISA and flow cytometry, respectively. U266 cell proliferation, viability and apoptosis were measured using BrdU, CCK-8 and Annexin V/propidium iodide assays, respectively. Total and phospo-(p-)JAK2 and p-STAT3 levels were measured by western blotting. Dual-luciferase reporter assays were performed to validate the predicted target binding sites. miR-451a expression was low in patients with MM and was associated with the Revised International Staging System (R-ISS) stage. IL-6 concentrations were significantly higher in patients with MM than in normal controls and were inversely associated with miR-451a levels (r=-0.96, P<0.0001). IL-6R levels were positively correlated with the R-ISS stage. miR-451a was downregulated, and IL-6R was upregulated in myeloma cell lines. Treatment with an miR-451a mimic inhibited viability and induced apoptosis in U266 cells. p-JAK2 and p-STAT3 levels were significantly lower in mimic-treated U266 cells than in control cells. Thus, miR-451a was shown to regulate myeloma cell proliferation and apoptosis via the IL-6R/JAK2/STAT3 pathway and may be used to predict patient prognosis.
由白细胞介素 -6（IL -6）介导的IL -6R/JAK2/STAT3信号通路在多发性骨髓瘤（MM）的发生发展中起重要作用，这与微小RNA -451a（miR -451a）水平降低有关。然而，miR -451a在MM中的生物学功能仍不清楚。采集MM患者的骨髓（BM），富集浆细胞。分别采用逆转录定量PCR、酶联免疫吸附测定（ELISA）和流式细胞术评估BM中miR -451a、IL -6和IL -6R水平以及Ki -67表达强度。分别采用BrdU、CCK -8和Annexin V/碘化丙啶检测法测定U266细胞增殖、活力和凋亡情况。通过蛋白质免疫印迹法检测总JAK2和磷酸化（p -）JAK2以及p -STAT3水平。进行双荧光素酶报告基因检测以验证预测的靶标结合位点。miR -451a在MM患者中表达较低，且与修订的国际分期系统（R -ISS）分期相关。MM患者的IL -6浓度显著高于正常对照，且与miR -451a水平呈负相关（r = -0.96，P <0.0001）。IL -6R水平与R -ISS分期呈正相关。在骨髓瘤细胞系中，miR -451a表达下调，而IL -6R表达上调。用miR -451a模拟物处理可抑制U266细胞活力并诱导其凋亡。模拟物处理的U266细胞中p -JAK2和p -STAT3水平显著低于对照细胞。因此，miR -451a被证明可通过IL -6R/JAK2/STAT3信号通路调节骨髓瘤细胞增殖和凋亡，并且可能用于预测患者预后。
