陕西医学杂志
陝西醫學雜誌
협서의학잡지
Shaanxi Medical Journal
2015年
10期
1275-1277
,共3页
雷秦%王梦昌%王娟
雷秦%王夢昌%王娟
뢰진%왕몽창%왕연
苦参碱%白血病,早幼粒细胞,急性%受体,自然杀伤细胞%HL60细胞%NKG2D配体
苦參堿%白血病,早幼粒細胞,急性%受體,自然殺傷細胞%HL60細胞%NKG2D配體
고삼감%백혈병,조유립세포,급성%수체,자연살상세포%HL60세포%NKG2D배체
Matrine%Leukemia,promyelocytic,acute%Receptors,naturalkillercell%HL60%NKG2D ligands
目的：探讨苦参碱对N K细胞杀伤急性早幼粒细胞白血病细胞株 H L60的影响及作用机制。方法：实时定量PCR（qRT‐PCR）检测苦参碱处理前后 HL60细胞表面NK细胞活化性受体凝集素样同型二聚体（NKG2D）配体分子MICA、MICB、ULBP1、ULBP2和 ULBP3表达情况， CFSE染色流式法检测苦参碱作用前后 HL60细胞对 NK 细胞杀伤敏感性的改变。结果：qRT‐PCR结果表明1．0 mg／ml苦参碱处理HL60细胞24h后HL60细胞表面MICA和ULBP2表达水平较正常对照组分别增加2．34倍和2．86倍，而MICB、ULBP1和ULBP3与处理前相比差异无统计学意义（P＞0．05）；处理48h后HL60细胞表面MICA／B、ULBP1／2／3表达水平较处理前均有不同程度升高，尤以ULBP2和 ULBP3升高最明显，较正常对照组分别增加了3．74倍和3．22倍。CFSE染色流式数据显示效靶比例5∶1和10∶1条件下，苦参碱均可加强NK细胞对 HL60细胞的杀伤效率。结论：苦参碱可促进 HL60细胞对NK细胞杀伤敏感性，其机制与诱导 HL60细胞表面NKG2D配体表达上调密切相关。
目的：探討苦參堿對N K細胞殺傷急性早幼粒細胞白血病細胞株 H L60的影響及作用機製。方法：實時定量PCR（qRT‐PCR）檢測苦參堿處理前後 HL60細胞錶麵NK細胞活化性受體凝集素樣同型二聚體（NKG2D）配體分子MICA、MICB、ULBP1、ULBP2和 ULBP3錶達情況， CFSE染色流式法檢測苦參堿作用前後 HL60細胞對 NK 細胞殺傷敏感性的改變。結果：qRT‐PCR結果錶明1．0 mg／ml苦參堿處理HL60細胞24h後HL60細胞錶麵MICA和ULBP2錶達水平較正常對照組分彆增加2．34倍和2．86倍，而MICB、ULBP1和ULBP3與處理前相比差異無統計學意義（P＞0．05）；處理48h後HL60細胞錶麵MICA／B、ULBP1／2／3錶達水平較處理前均有不同程度升高，尤以ULBP2和 ULBP3升高最明顯，較正常對照組分彆增加瞭3．74倍和3．22倍。CFSE染色流式數據顯示效靶比例5∶1和10∶1條件下，苦參堿均可加彊NK細胞對 HL60細胞的殺傷效率。結論：苦參堿可促進 HL60細胞對NK細胞殺傷敏感性，其機製與誘導 HL60細胞錶麵NKG2D配體錶達上調密切相關。
목적：탐토고삼감대N K세포살상급성조유립세포백혈병세포주 H L60적영향급작용궤제。방법：실시정량PCR（qRT‐PCR）검측고삼감처리전후 HL60세포표면NK세포활화성수체응집소양동형이취체（NKG2D）배체분자MICA、MICB、ULBP1、ULBP2화 ULBP3표체정황， CFSE염색류식법검측고삼감작용전후 HL60세포대 NK 세포살상민감성적개변。결과：qRT‐PCR결과표명1．0 mg／ml고삼감처리HL60세포24h후HL60세포표면MICA화ULBP2표체수평교정상대조조분별증가2．34배화2．86배，이MICB、ULBP1화ULBP3여처리전상비차이무통계학의의（P＞0．05）；처리48h후HL60세포표면MICA／B、ULBP1／2／3표체수평교처리전균유불동정도승고，우이ULBP2화 ULBP3승고최명현，교정상대조조분별증가료3．74배화3．22배。CFSE염색류식수거현시효파비례5∶1화10∶1조건하，고삼감균가가강NK세포대 HL60세포적살상효솔。결론：고삼감가촉진 HL60세포대NK세포살상민감성，기궤제여유도 HL60세포표면NKG2D배체표체상조밀절상관。
Objective:To explore the influence of killing efficiency of natural killer cell(NK)on HL60 cell line by matrine and investigate its underlying molecular mechanism .Methods:The expression of NKG2D ligands (major histocompatibility complex class I chain‐related molecule A or B (MICA/B) ,UL16‐binding proteins(ULBP) 1 ,2 ,and 3 on HL60 cells were analyzed before and after treated with matrine by quantitive real‐time PCR (qRT‐PCR) .The cytotoxic sensitivity of HL60 to NK cell was detected by FCM after CFSE staining at different effect‐to‐target (E/T) cell ratios .Results :After treatment with matrine for 24h ,MICA and ULBP2 expression on HL60 cells were significantly increased ,however ,there was no statistically significant difference between MICB ,ULBP1/3 ex‐pression and normal control group .After treatment with matrine for 48h ,the expression of MICA/B ,ULBP1/2/3 were increased inordinately .Flow cytometry results showed that at the ratio of E/T with whether 5∶1 or 10∶1 ,the proportion of the killed HL60 cells were increased by matrine treatment .Conclusion :Matrine can promote the killing efficiency of NK on HL60 cell line ,which is closely related to up‐regulation of NKG2D ligands .
