苦参碱通过诱导NKG2D配体高表达增强NK细胞对ABCG2high耐药鼻咽癌细胞杀伤活性

目的 探讨苦参碱提高ABCG_2~(High)三磷酸腺苷(ATP)结合转运蛋白G超家族成员2]耐药鼻咽癌细胞对Allo-NK细胞杀伤敏感性的机制.方法 利用免疫磁珠技术分离ABCG_2~(High)CNE2/DDP细胞及Allo-NK细胞,流式细胞技术检测分离后细胞纯度及经苦参碱处理前后靶细胞NKG2D配体表达率,LDH释放测定法检测经苦参碱处理前后ABCG_2~(High)CNE2/DDP细胞对Allo-NK细胞的杀伤敏感性.结果 ABCG_2~(High)CNE2/DDP细胞分离后ABCG_2表达率为(91.40±2.32)%,分选后NK细胞CD3~-CD16~+CD56~+细胞的纯度达90%以上,经苦参碱处理之后靶细胞MICA、MICB、ULBP1、ULBP2、ULBP3表达率,由药物处理之前的(2.92±0.33)%、(4.27±0.33)%、(5.80±0.62)%、(11.10±3.15)%、(7.75±1.14)%分别上升到(11.30±0.89)%、(14.29±2.61)%、(12.56±1.06)%、(43.24±4.43)%、(12.77±1.06)%.在效靶比为10:1、20:1,Allo-NK细胞对苦参碱处理前后ABCG2HighCNE2/DDP细胞的杀伤率分别为(15.32±1.34)%、(27.26±6.81)%及(28.53±1.37)%、(42.72±2.80)%.处理前后杀伤率有显著性差异(F=29.05,P=0.000).结论 苦参碱通过诱导肿瘤细胞高表达NKG2D配体(MICA/B、ULBP1-3),使肿瘤细胞对Allo-NK细胞的杀伤敏感性增强.

Abstract：

Objective To investigate the mechanism underlying the effects of matrine in enhancing the cytotoxic sensitivity of CNE2/DDP cells highly expressing ATP-binding cassette superfamily G member 2(ABCG_2~(High) to allogenic natural killer (Allo-NK、cells.Methods ABCGr2~(High) CNE2/DDP cells and Allo-NK cells were isolated by magnetic activated eell sorting (MACS).Flow cytometry was used to evaluffte the purity of the isolated cells and the expression of NKG2D ligands on thetarget cells before and after incubation with matrine.The cytotoxic sensitivity of the treated and non.treated ABCG_2~(High)CNE2/DDP cells to Allo-NK cells was measured by LDH releasing assay. Resuits The expression rate of ABCG2 was (9 1.40±.-321%in ABCG_2~(High),CNE2/DDP cells. More than 90% of the isolated NK cells were identified to be CD3~-cD16~+CD56~+ cells.The expression rates of MICA,MICB,ULBP1,ULBP2,and ULBP3 on the target cells incubated with matrine increased from(2.92±0.33)%,(4.27±0.33)%,(5.80±0.62)%,(11.10±3.15)%,and(7.75±1.14)% to(11.30±0.89)%,(14.29±2.61)%,(12.56±1.06)%,(43.24±4.43)%,and(12.77±1.06)%,respectively.At the E:T ratio of 10:1 and 20:1,the cytotoxic sensitivity of ABCG2High cells to Allo-NK cells increased from(15.32±1.34)% and (27.26±6.81)% in un-treated cells to(28.53±1.37)% and(42.72±2.80)%in matrine-treated cells,respectively,showing significant differences in the cytotoxic sensitivity of the target cells in both groups produced by matrine treatment(F=29.05,P=0.000).Conclusion Matrine can up-regulate the expressions of NKG2D ligands (MICA/B and ULBP 1-3)in ABCG_2~(high) nasopharyngeal carcinoma cells,which results in increased cytotoxic sensitivity of the cells to Allo-NK cells.

Matrine enhances the cytotoxic sensitivity of ABCG2high nasopharyngeal carcinoma cells to natural killer cells by inducing high expression of NKG2D receptor ligands

Objective To investigate the mechanism underlying the effects of matrine in enhancing the cytotoxic sensitivity of CNE2/DDP cells highly expressing ATP-binding cassette superfamily G member 2(ABCG2High) to allogenic natural killer(Allo-NK) cells.Methods ABCG2High CNE2/DDP cells and Allo-NK cells were isolated by magnetic activated cell sorting(MACS).Flow cytometry was used to evaluate the purity of the isolated cells and the expression of NKG2D ligands on the target cells before and after incubation with ma...