两种门冬酰胺酶对Jurkat和RS4;11细胞的细胞毒性差异研究

The difference cytotoxicity of two asparaginases on Jurkat cells and RS4;11 cells

目的比较谷氨酰胺酶活性不同的两种门冬酰胺酶(L-ASP)对急性淋巴细胞白血病细胞株Jurkat(L-ASP耐药)和RS4;11(L-ASP敏感)的细胞毒性,探讨门冬酰胺酶的谷氨酰胺酶活性对抗肿瘤效应的影响。方法(1)通过基因工程方法获得野生型门冬酰胺酶(WT)和低谷氨酰胺酶活性的突变型门冬酰胺酶(MT)。(2)用CCK-8试剂盒检测WT和MT对Jukat细胞和RS4;11细胞的增殖毒性。(3)绘制WT和MT作用下Jurkat细胞的生长曲线。(4)用流式细胞术检测WT和MT诱导Jurkat细胞凋亡情况。结果(1)与WT相比,MT的谷氨酰胺酶活性和门冬酰胺酶活性比值明显降低(0.9%±0.1%vs.12.2%±2.8%,P<0.05)。(2)1.0 IU/mL的WT和MT处理Jurkat细胞48h抑制率分别为(96.07±1.19)%和(45.60±4.38)%,差异具有显著性(P<0.05);WT和MT对RS4;11细胞的半数抑制浓度(IC50)分别为(0.127±0.012)IU/L和(0.116±0.014)IU/L,差异无显著性(P=0.331)。(3)生长曲线显示0.1 IU/mL的WT显著抑制Jurkat细胞增殖,0.1 IU/mL的MT不能抑制Jurkat细胞增殖。(4)1.0 IU/mL的WT和MT处理Jurkat细胞24h细胞凋亡率分别为(37.27±6.89)%和(13.02±3.87)%,差异具有显著性(P<0.05)。结论L-ASP的谷氨酰胺酶活性可以增强L-ASP对Jurkat细胞的抗肿瘤效应,但不能增强L-ASP对RS4;11细胞的抗肿瘤效应。

Objective To compare the cytotoxicity of two L-Asparaginases(L-ASP)with different glutaminase activities on acute lymphoblastic leukemia cell lines,Jurkat(L-ASP resistant)and RS4;11(L-ASP sensitive),and to explore whether the glutaminase activity of L-ASP could enhance the anticancer efficacy.Methods(1)Mutant asparaginase(MT)with low glutaminase activity and Wild type asparaginase(WT)were obtained by genetic engineering.(2)CCK-8 kit was used to detect the proliferation toxicity of WT and MT on Jukat and RS4;11.(3)The cell growth curves of Jurkat cells treated with WT or MT were drawn.(4)Flow cytometry was used to measure the apoptosis of Jurkat cells induced by WT and MT.Results(1)The glutaminase activity of MT was lower than that of WT(0.9%±0.1%vs.12.2%±2.8%,P<0.05).(2)The inhibition rate of Jurkat cells treated with 1.0 IU/mL WT for 48h was significantly higher than with MT,respectively(96.07%±1.19%and 45.60%±4.38%,P<0.05).(3)The cell growth curves showed that 0.1 IU/mL WT inhibits the proliferation of Jurkat cells,while 0.1 IU/mL MT could not prevent Jurkat cells from proliferating.(4)The apoptosis rates of Jurkat cells treated with WT and MT at 1.0 IU/mL for 24h were(37.27±6.89)%and(13.02±3.87)%respectively,with significant difference(P<0.05).Conclusions In vitro,the glutaminase activity of L-ASP could enhance the anticancer efficacy of L-ASP on Jurkat cells,but not on RS4;11 cells.