Study On The Effect And Mechanism Of Tocilizumab On Proliferation And Apoptosis Of Multiple Myeloma Cells

Objective: To investigate the effect and possible mechanism of IL-6 receptor antibody(tocilizumab)on proliferation and apoptosis of multiple myeloma cells.Methods:Human myeloma cell line RPMI8226 cells were selected as the research object,and the minimum effective concentration of tocilizumab was determined by CCK-8 method after48 hours of treatment with different concentrations of tocilizumab.After 24 hours of treatment with low,medium and high concentrations of tocilizumab,the effects of RPMI8226 cells proliferation,apoptosis and JAK1 and STAT3 gene expression were detected by CCK-8 method,Annexin V-PE/7-AAD double staining method and q RT-PCR method.Results:1.The CCK-8 method determined that the minimum effective concentration of tocilizumab was 10μg/m L,and 10μg/m L,20μg/m L,50μg/m L tocilizumab were selected for subsequent studies.2.The results of RPMI8226 cells proliferation detected by CCK-8 showed that with the increase of tocilizumab concentration,the cell survival rate gradually decreased,and the cell survival rates of low,medium and high dose tocilizumab groups were 82.78%,75.38% and 66.09%,respectively,and the difference between the three groups was statistically significant(P<0.05).3.Flow cytometry Annexin V-PE/7-AAD double staining detection of apoptosis showed that tocilizumab could induce apoptosis in RPMI8226 cells,with the increase of tocilizumab concentration,the apoptosis rate increased,the difference was statistically significant(P<0.05),and the apoptosis rates of tocilizumab groups at low dose,medium and high doses were 12.68%,23.50% and29.13%,respectively.4.The expression results of JAK1 and STAT3 m RNA by q RT-PCR showed that tocilizumab could reduce the expression of JAK1 and STAT3 m RNA in myeloma cell line RPMI8226 cells,and the differences between different concentration groups were statistically significant(P<0.05).Conclusion: 1.Tocilizumab can inhibit the proliferation of human myeloma cell line RPMI8226 cells,promote apoptosis,and is concentration-dependent.2.Tocilizumab may affect myeloma cell proliferation and apoptosis by blocking the IL-6-mediated JAK1/STAT3 signaling pathway.