氧化苦参碱调控PD-L1对弥漫大B淋巴瘤细胞增殖的影响

Effects of oxymatrine on the proliferation of diffuse large B lymphoma cells by regulating PD-L1

目的探究氧化苦参碱对弥漫大B淋巴瘤细胞增殖、凋亡的影响及其分子机制。方法将人弥漫大B淋巴瘤细胞OCI-LY19随机分为对照组(正常培养)、低剂量实验组(25.00μmol·L^(-1)氧化苦参碱)、中剂量实验组(50.00μmol·L^(-1)氧化苦参碱)、高剂量实验组(100.00μmol·L^(-1)氧化苦参碱)、氧化苦参碱+si-NC组(转染si-NC+100.00μmol·L^(-1)氧化苦参碱)、氧化苦参碱+si-PD-L1组(转染si-PD-L1+100.00μmol·L^(-1)氧化苦参碱)、氧化苦参碱+Vector组(转染si-NC+100.00μmol·L^(-1)氧化苦参碱)、氧化苦参碱+PD-L1组[转染程序性死亡受体配体1(PD-L1)+100.00μmol·L^(-1)氧化苦参碱]。用5-乙炔基-2’-脱氧尿苷(EdU)实验检测细胞增殖情况,用蛋白质印迹法检测蛋白表达水平,用流式细胞术实验检测细胞凋亡情况。结果对照组、高剂量实验组、氧化苦参碱+si-NC组、氧化苦参碱+si-PD-L1组、氧化苦参碱+Vector组、氧化苦参碱+PD-L1组的细胞增殖率分别为(33.88±2.79)%、(15.23±1.32)%、(15.84±1.58)%、(10.13±0.90)%、(16.14±1.20)%和(20.84±1.88)%,周期蛋白依赖性激酶4(CDK4)蛋白相对表达水平分别为0.92±0.11、0.37±0.04、0.35±0.04、0.24±0.03、0.38±0.06和0.71±0.06,细胞凋亡率分别为(3.20±0.07)%、(25.35±2.01)%、(24.81±1.91)%、(30.27±1.65)%、(24.39±2.73)%和(17.97±1.27)%,高剂量实验组的上述指标与对照组比较,在统计学上差异均有统计学意义(均P<0.05);氧化苦参碱+si-PD-L1组的上述指标与氧化苦参碱+si-NC组比较,在统计学上差异均有统计学意义(均P<0.05);氧化苦参碱+PD-L1组的上述指标与氧化苦参碱+Vector组比较,在统计学上差异均有统计学意义(均P<0.05)。结论氧化苦参碱可通过下调PD-L1表达抑制OCI-LY19细胞增殖,并诱导凋亡;PD-L1可能是氧化苦参碱治疗弥漫大B淋巴瘤的潜在靶点。

Objective To investigate the effects of oxymatrine on proliferation and apoptosis of diffuse large B-cell lymphoma and its molecular mechanism.Methods Human diffuse large B lymphoma cells OCI-LY19 were randomly divided into control group(normal culture),experimental-L group(25.00μmol·L^(-1)oxymatrine),experimental-M group(50.00μmol·L^(-1)oxymatrine),experimental-H group(100.00μmol·L^(-1)oxymatrine),Oxymatrine+si-NC group(transfected with si-NC+100.00μmol·L^(-1)oxymatrine),Oxymatrine+si-P D-L1 group[transfected with si-programmed death receptor ligand 1(PD-L1)+100.00μmol·L^(-1)oxymatrine],Oxymatrine+Vector group(transfected with si-NC+100.00μmol·L^(-1)oxymatrine)and oxymatrine+PD-L1 group(transfected with PD-L1+100.00μmol·L^(-1)oxymatrine).5-acetylidene-2′-deoxyuridine(EdU)assay was used to detect cell proliferation;Western blot assay was used to detect protein expression;flow cytometry assay was used to detect cell apoptosis.Results The EdU cell proliferation rates in control group,experimental-H group,oxymatrine+si-NC group,oxymatrine+si-PD-L1 group,oxymatrine+Vector group and oxymatrine+PD-L1 group were(33.88±2.79)%,(15.23±1.32)%,(15.84±1.58)%,(10.13±0.90)%,(16.14±1.20)%and(20.84±1.88)%,respectively;the expressions of Cyclin-dependent kinase 4(CDK4)protein were 0.92±0.11,0.37±0.04,0.35±0.04,0.24±0.03,0.38±0.06 and 0.71±0.06,respectively;the apoptosis rates were(3.20±0.07)%,(25.35±2.01)%,(24.81±1.91)%,(30.27±1.65)%,(24.39±2.73)%and(17.97±1.27)%,respectively.The above indicators:Experimental-H group were compared with the control group respectively,and the differences were statistically significant(all P<0.05);there were significant differences between oxymatrine+si-PD-L1 group and oxymatrine+si-NC group(all P<0.05);there were significant differences between oxymatrine+PD-L1 group and oxymatrine+Vector group(all P<0.05).Conclusion Oxymatrine can inhibit the proliferation of OCI-LY19 cells and induce apoptosis by down-regulating the expression of PD-L1.PD-L1 may be a potential target of oxymatrine in the treatment of diffuse large B lymphoma.