Effects of NEK2 knockdown on multiple myeloma cell-mediated bone destruction and NF-κB signaling pathway
WANG Peng
GUO Jie
LU Laitong
ZHONG Jiangbo
PAN Pan
WANG Hongchao
LIU Jing
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.
Keywords：multiple myelomamyeloma bone diseasebone destructioncentrosome-associated kinase 2nucle-ar factor-κB signaling pathwaymultiple myeloma cellsbone marrow stromal cells
Publication Date：2025-05-25
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：5( 7-11 )