Apoptotic Effect Of Pomalidomide On MM1.S In Multiple Myeloma Cells And Its Regulation On Cereblon Expression

Objective: To explore the effect of pomalidomide on proliferation and apoptosis of human multiple myeloma cell line MM1.S and its effect on Cereblon expression,and further explore its regulation on key enzymes and key proteins of glucose metabolism.Methods: Different concentrations of pomalidomide acted on MM1.S cells for 24,48,and 72 hours,and the inhibition rate of MM1.S cells was detected by CCK-8method.Apoptosis rate of MM1.S cells detected by Annexin V-FITC/PI double staining.Real-time quantitative PCR detection of CRBN gene expression levels.Western blot analysis of the effect of pomalidomide on the expression of CRBN protein,glucose metabolism key protein AMPK and p-AMPK in MM1.S cells.Results: 1.Pomalidomide has an inhibitory effect on MM1.S cells,and its proliferation inhibition is time-and dose-dependent(r=0.981,r=0.990).At 72 h,the inhibition rate of cell proliferation in 40 and 80 μmol/L group was P<0.001,the difference was statistically significant.2.Pomalidomide induces apoptosis in MM1.S cells.The concentrations of pomalidomide were 0,40 and 80 μmol/L for 72 h,respectively.The apoptotic rates were 0.003 ± 0.001,8.333 ± 0.260,10.730 ± 0.210.The higher the drug concentration,the higher the apoptotic rate(r=0.952),the difference is statistically significant.3.When the concentration of pomalidomide was 0,40 and 80 μmol/L,the expression of CRBN gene after treatment for MM1.S cells for 72 hours was1.487±0.340,0.478±0.076,0.211±0.054,0.055±0.005.In a dose-dependent manner,the expression of CRBN gene was down-regulated with increasing drug concentration(P<0.05),and the difference was statistically significant.4.Compared with the control group,the expression of CRBN protein in the pomalidomide group(40 and 80 μmol/L)was significantly decreased,and the expression of CRBN protein decreased with increasing drug concentration.5.As the concentration of pomalidomide increases,the activities of key enzymes of glucose metabolism(pyruvate kinase,lactate dehydrogenase)and lactic acid content gradually decrease.6.Compared with the control group,with the increase of the concentration of pomalidomide,the expression of AMPK protein,a key enzyme of glucose metabolism,remained unchanged,and the expression of p-AMPK protein gradually increased.Conclusion: Pomalidomide can inhibit the proliferation of MM1.S cells and promote its apoptosis.A certain concentration of pomalidomide can reduce the expression of CRBN gene in MM1.S cells and down-regulate its protein expression.Pomalidomide can regulate the glucose metabolism pathway when it acts on MM1.S cells.