Effect of conditioned medium of diffuse large B-cell lymphoma cells on proliferation and apoptosis of human bone marrow mesenchymal stem cells

Yuan Xiaoshuang; Yang Xu; Yang Bo; Chen Xiaoxu; Tian Ting; Wang Feiqing; Li Yanju; Liu Yang; Yang Wenxiu

doi:10.12307/2026.524

Abstract：BACKGROUND:The relationship between bone marrow microenvironment with growth,survival,and drug resistance of diffuse large B-cell lymphoma has become a hot topic of research,but the effect of diffuse large B-cell lymphoma cell conditioned medium on bone marrow mesenchymal stem cells has not been reported. OBJECTIVE:To investigate the effect of cell conditioned medium of diffuse large B-cell lymphoma on proliferation and apoptosis of bone marrow mesenchymal stem cells. METHODS:Bone marrow mesenchymal stem cells were extracted from bone marrow blood of healthy donors by Ficoll density gradient centrifugation method and purified by adhesion method.Meanwhile,two kinds of diffuse large B-cell lymphoma cell supernatant(SU-DHL-2 and OCI-LY3)were used to prepare conditional culture medium.The cells were grouped according to different media:human bone marrow mesenchymal stem cells in the control group were cultured with L-DMEM only.The human bone marrow mesenchymal stem cells in the CM-SU-DHL-2 and CM-OCI-LY3 groups were cultured with 20%SU-DHL-2 cell conditioned culture medium or 20%OCI-LY3 cell conditioned culture medium and 80%L-DMEM complete culture medium.The proliferation of bone marrow mesenchymal stem cells was observed by CCK-8,EDU and crystal violet staining.The migration of bone marrow mesenchymal stem cells was evaluated by scratch method.The cell cycle and apoptosis of bone marrow mesenchymal stem cells were detected by flow cytometry.The expression levels of P21,P16,Bcl-2,and BTK mRNA and protein were detected by real-time PCR and western blot assay. RESULTS AND CONCLUSION:(1)Compared with the control group,the conditioned medium of SU-DHL-2 and OCI-LY3 cells could significantly promote the proliferation of bone marrow mesenchymal stem cells(P<0.05),which might be closely related to the low expression of P21 and P16 proteins(P<0.05).(2)Compared with the control group,the conditioned medium of SU-DHL-2 and OCI-LY3 cells significantly promoted the migration of bone marrow mesenchymal stem cells(P<0.05).(3)Compared with the control group,the conditioned medium of SU-DHL-2 and OCI-LY3 cells inhibited the apoptosis of bone marrow mesenchymal stem cells(P<0.05),which might be closely related to the high expression of Bcl-2 and BTK mRNA and protein(P<0.05).The results showed that the conditioned medium of diffuse large B-cell lymphoma cells could promote the proliferation and inhibit apoptosis of bone marrow mesenchymal stem cells.