Comparative Study On The Effects Of Triptolide And Thalidomide On Multiple Myeloma Cell Lines And Bone Marrow Mesenchymal Stem Cells

Objective To observe the expression differences of triptolide and thalidomide in multiple myeloma cell lines and bone marrow mesenchymal stem cells about cytokine, and mechanism discussion, to provide some experimental basis for clinical application.Methods 1.MTT assay was used to detect RPMI8226, ARP-1 (CCK8) and bone marrow mesenchymal stem cells proliferation; 2 Annexin V-FITC/PI double staining to detect cell apoptosis, and cell apoptosis was observed by Wright staining morphology; cell cycle analysis by flow cytometry; 3.RT-qPCR detection by Triptolide thalidomide effect on bone marrow mesenchymal stem cell factor IL-6 expression, IL-1 beta and SCF mRNA cells after 4 Western; Blot analysis by Triptolide and thalidomide on expression in bone marrow mesenchymal stem cells NF-κB/P65 protein level after 5 ELISA; enzyme linked immunosorbent assay after determination of triptolide, thalidomide on the role of bone marrow mesenchymal stem cells after cytokine VEGF expression.Results 1. MTT than colorimetric method (CCK8) detection results showed that triptolide can significantly inhibit the growth of RPMI8226 and ARP-1, in a concentration dependent manner.0-8ng/ml triptolide treatment of RPMI 8226 and ARP-1 cells for 72h and compared with the control group, P< 0.001, with an IC50 of 118.7±0.08ng/ml and 0.457±0.13ng/ml. RPMI8226, ARP-172h after thalidomide treatment, compared with control group P>0.05, no statistical significance.2 induced apoptosis in RPMI8226 and ARP-1 cells in a dose dependent manner and accompanied by typical morphological changes of cells. The cell cycle of RPMI8226 and ARP-1 in the S phase of the cell cycle was. Compared. The triptolide and thalidomide treatment after bone marrow mesenchymal stem cells by qPCR assay, cell supernatant IL-6, IL-1 beta, the SCF expression decreased, compared with the blank control group, triptolide (P<0.05), thalidomide group only IL-6 and blank control group (P<0.05).4. By Western blot after triptolide treatment after bone marrow mesenchymal stem cells can decreased the expression of NF-κB/p65, P< 0.05, thalidomide group no significant difference (P>0.05). Detection of 5.ELISA after triptolide treatment after bone marrow mesenchymal stem cells to express VEGF was significantly lower than that in the thalidomide group, compared with the blank control group, triptolide (P<0.05), statistical significance. Thalidomide group P>0.05, no statistical significance.Conclusion 1. TPL can directly inhibit the growth of myeloma cells RPMI8226 and ARP-1 and BMMSCs, in a concentration dependent manner, the MM cells significantly inhibited the proliferation of strong in BMMSCs; 2, the TPL can be induced MM cell line RPMI8226 and ARP-1 and BMMSCs apoptosis was concentration dependent, in MM cells induced apoptosis is significantly stronger in BMMSCs; 3, TPL can directly kill MM cells. The BMMSCs could possible does not have a direct killing effect, but can affect its differentiation and expression; reduce the expression of inflammation mediators IL-6, IL-1, SCF, VEGF, improve the bone marrow hematopoietic microenvironment, further inhibiting proliferation of MM cells.4, TPL may induce apoptosis by inhibiting the activation of NF-kappa B/P65, and inhibit the proliferation of tumor cells.