山东医药2025，Vol.65Issue(5)：7-11,5.DOI:10.3969/j.issn.1002-266X.2025.05.002
NEK2基因敲低对多发性骨髓瘤细胞介导骨质破坏及NF-κB信号通路的影响
Effects of NEK2 knockdown on multiple myeloma cell-mediated bone destruction and NF-κB signaling pathway
摘要
Abstract
Objective To investigate the effects of centrosome-associated kinase 2(NEK2)gene knockdown on multiple myeloma(MM)cell-mediated bone destruction and the nuclear factor kappa B(NF-κB)signaling pathway.Methods MM cell lines(MM.1S cells and RPMI 8226 cells)were used,with both NEK2 knockdown group and negative control group established for each.Lentiviral vectors carrying NEK2-RNA interference(shRNA)or control virus(CON077)were used for transfection.NEK2 mRNA expression was detected by real-time reverse transcription quantita-tive polymerase chain reaction(RT-qPCR)to confirm transfection efficiency.Bone marrow stromal cells(BMSCs)were di-vided into the negative control group and NEK2 knockdown group,which were co-cultured with RPMI 8226 cells or NEK2-knockdown RPMI 8226 cells.RT-qPCR was used to measure the mRNA levels of bone destruction-related markers,including receptor activator of NF-κB ligand(RANKL)and osteoprotegerin(OPG).MM cells from the NEK2 knockdown group and negative control group were selected.Cell proliferation(absorbance value)was assessed using the CCK-8 assay,apoptosis rates were measured by flow cytometry,and NF-κB pathway-related proteins[NF-κB,heparanase(HPSE)]and their mRNA levels were detected by Western blotting and RT-qPCR,respectively.Results NEK2 mRNA expression was significantly lower in the NEK2 knockdown groups of MM.1S and RPMI 8226 cells than in the negative control groups(both P<0.05),with knockdown efficiencies of 92.2%and 64.0%,respectively.Compared with the negative control group of BMSCs,the NEK2 knockdown group had lower RANKL mRNA expression(P<0.05),while OPG mRNA expres-sion was higher;no statistically significant difference was found(P>0.05).Compared with the negative control groups of MM.1S cells and RPMI-8226 cells,the NEK2 knockdown group of MM.1S and RPMI 8226 cells had lower absorbance val-ues,higher apoptosis rates,lower NF-κB and HPSE mRNA levels,and higher NF-κB and HPSE protein expression levels(all P<0.05).Conclusion NEK2 knockdown may inhibit MM cell proliferation,promote apoptosis,and up-regulate HPSE expression by modulating the NF-κB signaling pathway,thereby suppressing MM cell-mediated bone destruction.关键词
多发性骨髓瘤/骨髓瘤骨病/骨质破坏/中心体相关激酶2/核因子κB信号通路/多发性骨髓瘤细胞/骨髓基质细胞Key words
multiple myeloma/myeloma bone disease/bone destruction/centrosome-associated kinase 2/nucle-ar factor-κB signaling pathway/multiple myeloma cells/bone marrow stromal cells分类
临床医学引用本文复制引用
王鹏,郭洁,路来同,仲江波,潘盼,王洪超,刘静..NEK2基因敲低对多发性骨髓瘤细胞介导骨质破坏及NF-κB信号通路的影响[J].山东医药,2025,65(5):7-11,5.基金项目
山东省自然科学基金青年项目(ZR2020QH121) （ZR2020QH121）
山东省医药卫生科技发展计划项目(2018WS481). （2018WS481）
