CAJ | 학술논문

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.
목적 탐토고삼감제고ABCG_2~(High)[삼린산선감(ATP)결합전운단백G초가족성원2]내약비인암세포대Allo-NK세포살상민감성적궤제.방법 이용면역자주기술분리ABCG_2~(High)CNE2/DDP세포급Allo-NK세포,류식세포기술검측분리후세포순도급경고삼감처리전후파세포NKG2D배체표체솔,LDH석방측정법검측경고삼감처리전후ABCG_2~(High)CNE2/DDP세포대Allo-NK세포적살상민감성.결과 ABCG_2~(High)CNE2/DDP세포분리후ABCG_2표체솔위(91.40±2.32)%,분선후NK세포CD3~-CD16~+CD56~+세포적순도체90%이상,경고삼감처리지후파세포MICA、MICB、ULBP1、ULBP2、ULBP3표체솔,유약물처리지전적(2.92±0.33)%、(4.27±0.33)%、(5.80±0.62)%、(11.10±3.15)%、(7.75±1.14)%분별상승도(11.30±0.89)%、(14.29±2.61)%、(12.56±1.06)%、(43.24±4.43)%、(12.77±1.06)%.재효파비위10:1、20:1,Allo-NK세포대고삼감처리전후ABCG2HighCNE2/DDP세포적살상솔분별위(15.32±1.34)%、(27.26±6.81)%급(28.53±1.37)%、(42.72±2.80)%.처리전후살상솔유현저성차이(F=29.05,P=0.000).결론 고삼감통과유도종류세포고표체NKG2D배체(MICA/B、ULBP1-3),사종류세포대Allo-NK세포적살상민감성증강.
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.