Jin Yueling, Dai Zhensheng
Department of Pathology, Shanghai University of Medicine and Health Sciences, Shanghai 200237, China.
Department of Hematology, Shanghai Pudong Hospital Affiliated to Fudan University, Shanghai 201399, China.
Biomed Pharmacother. 2016 Mar;78:264-271. doi: 10.1016/j.biopha.2016.01.012. Epub 2016 Feb 2.
This study aimed to explore the influence of USO1 on multiple myeloma (MM) cell proliferation and apoptosis and the related molecular mechanism.
The expression of USO1 and MIF in MM tissues and cells, normal bone marrow tissues and cells were determined by qRT-PCR and western blot assay. The cell proliferation and apoptosis of MM cells before and after knockdown of USO1 were determined by MTT assay and flow cytometry, respectively. Before and after knockdown of USO1, the expression of the proliferation-related genes cyclin D1, Mcm2 and PCNA in MM cells was determined by qRT-PCR and western blot assay. The protein level of p-Erk1/2 and MIF was determined by western blot assay and ELISA, respectively.
The expression levels of USO1 and MIF in MM tissues and cells were much higher than those in normal bone marrow tissues and cells. Knockdown of USO1 resulted in the inhibited ability of cell proliferation and induced cell apoptosis. The expression of cyclin D1, Mcm2, PCNA and p-Erk1/2 decreased significantly after knockdown of USO1 as well as the decreased MIF secretion.
USO1 gene may be a promising target for the therapy of human MM and its diagnosis marker.
本研究旨在探讨USO1对多发性骨髓瘤（MM）细胞增殖和凋亡的影响及其相关分子机制。
采用qRT-PCR和蛋白质免疫印迹法检测MM组织和细胞、正常骨髓组织和细胞中USO1和MIF的表达。分别采用MTT法和流式细胞术检测敲低USO1前后MM细胞的增殖和凋亡情况。敲低USO1前后，采用qRT-PCR和蛋白质免疫印迹法检测MM细胞中增殖相关基因细胞周期蛋白D1、微小染色体维持蛋白2（Mcm2）和增殖细胞核抗原（PCNA）的表达。分别采用蛋白质免疫印迹法和酶联免疫吸附测定法检测p-Erk1/2和MIF的蛋白水平。
MM组织和细胞中USO1和MIF的表达水平明显高于正常骨髓组织和细胞。敲低USO1导致细胞增殖能力受到抑制并诱导细胞凋亡。敲低USO1后，细胞周期蛋白D1、Mcm2、PCNA和p-Erk1/2的表达以及MIF分泌均显著降低。
USO1基因可能是人类MM治疗的一个有前景的靶点及其诊断标志物。
