Li Xiaoya, Ding Lingzhi, Gu Geyu, Zheng Changjun, Pan Chenshuai, Zheng Qi, Xiang Ting
Department of Orthopedic, Taizhou Central Hospital (Taizhou University Hospital), Taizhou, Zhejiang 318000, China.
Department of Nutrition, Taizhou First people's Hospital, Taizhou, Zhejiang 318020, China.
Evid Based Complement Alternat Med. 2021 Sep 13;2021:4630934. doi: 10.1155/2021/4630934. eCollection 2021.
This study aims to explore circ_0058063 effect on multiple myeloma cells malignant phenotype and its feasible mechanism.
We selected 47 cases of multiple myeloma tissues and 47 cases of normal bone marrow tissues and then used RT-qPCR method to test circ_0058063 and miR-635 expression in the tissues. Myeloma cells RPMI8226 were transfected with si-circ_0058063, miR-635 mimic, and si-circ_0058063 + anti-miR-635, respectively. Then, we adopt CCK-8 method, flow cytometry method, and Transwell and western blot methods to detect the influences of knockdown of circ_0058063 or miR-635 overexpression on RPMI8226 cell proliferation, apoptosis, migration, and invasion and also Ki-67, Bax, Bcl-2, MMP-2, and MMP-9 protein expression. The dual luciferase reporter gene assay experiment proved that it has regulatory relationship between circ_0058063 and miR-635.
circ_0058063 expression of multiple myeloma was higher than that in normal bone marrow tissue ( < 0.05), while miR-635 expression was lower than that in normal bone marrow tissue ( < 0.05). Knockdown of circ_0058063 or overexpression of miR-635 could reduce proliferation capacity, migration, invasion cell quantities, and Ki-67, MMP-2, MMP-9, and Bcl-2 protein expression ( < 0.05), while increasing apoptosis rate together with Bax protein expression ( < 0.05). circ_0058063 targets to negatively regulate miR-635, while knocking down miR-635 reverses the influences of knocking down circ_0058063 on RPMI8226 proliferation, apoptosis, migration, and invasion.
circ_0058063 expression increased in multiple myeloma tissues. Knocking down its expression may inhibit myeloma proliferation, migration, and invasion by targeting and upregulating miR-635 and also promote cell apoptosis. As for multiple myeloma treatment, circ_0058063/miR-635 may provide new molecular targets.
本研究旨在探讨circ_0058063对多发性骨髓瘤细胞恶性表型的影响及其可能机制。
选取47例多发性骨髓瘤组织和47例正常骨髓组织，采用RT-qPCR法检测组织中circ_0058063和miR-635的表达。分别用si-circ_0058063、miR-635模拟物、si-circ_0058063 + 抗miR-635转染骨髓瘤细胞RPMI8226。然后，采用CCK-8法、流式细胞术、Transwell和蛋白质印迹法检测敲低circ_0058063或过表达miR-635对RPMI8226细胞增殖、凋亡、迁移和侵袭以及Ki-67、Bax、Bcl-2、MMP-2和MMP-9蛋白表达的影响。双荧光素酶报告基因检测实验证实circ_0058063与miR-635之间存在调控关系。
多发性骨髓瘤中circ_0058063表达高于正常骨髓组织（<0.05），而miR-635表达低于正常骨髓组织（<0.05）。敲低circ_0058063或过表达miR-635可降低增殖能力、迁移和侵袭细胞数量以及Ki-67、MMP-2、MMP-9和Bcl-2蛋白表达（<0.05），同时增加凋亡率和Bax蛋白表达（<0.05）。circ_0058063靶向负调控miR-635，敲低miR-635可逆转敲低circ_0058063对RPMI8226增殖、凋亡、迁移和侵袭的影响。
多发性骨髓瘤组织中circ_0058063表达升高。敲低其表达可能通过靶向并上调miR-635抑制骨髓瘤的增殖、迁移和侵袭，还能促进细胞凋亡。对于多发性骨髓瘤的治疗，circ_0058063/miR-635可能提供新的分子靶点。
