中华神经医学杂志
中華神經醫學雜誌
중화신경의학잡지
CHINESE JOURNAL OF NEUROMEDICINE
2014年
4期
330-335
,共6页
神经胶质瘤%U251细胞%代谢型谷氨酸受体5亚型%增殖
神經膠質瘤%U251細胞%代謝型穀氨痠受體5亞型%增殖
신경효질류%U251세포%대사형곡안산수체5아형%증식
Glioma%U251 cell%Metabotropic glutamate receptor 5 subtype%Proliferation
目的 通过观察代谢型谷氨酸受体5亚型(mGluR5)的特异性拮抗剂2-甲基-6-苯基乙炔嘧啶(MPEP)对人胶质瘤细胞U251的影响间接探讨mGluR5对U251增殖的影响并对其机制进行分析.方法 体外常规培养人胶质瘤细胞株U251,噻唑蓝(MTT)检测100 μmol/L MPEP、20μmol/LSP600125(JNK特异性拮抗剂)、400感染复数(MOI) Ad-△ 169(c-Jun负显性病毒)作用48 h后U251细胞增殖的变化;Western blotting检测5、10、20、50、100 μmol/LMPEP作用24 h及100μmol/L MPEP作用4、8、12、24 h后U251细胞表达磷酸化c-Jun及磷酸化JNK的变化.结果 1 00 μmol/L MPEP、20 μmol/LSP600125、400MOIAd-△169处理48 h后U251细胞的吸光度值均明显低于对照组,差异有统计学意义(P＜0.05);5、10、20、50、100 μmol/L MPEP处理人胶质瘤细胞U251 24 h后的磷酸化c-Jun、磷酸化JNK表达量均明显低于对照组,而且随着用药浓度的增加,U251细胞磷酸化c-Jun、磷酸化JNK的表达越低,差异有统计学意义(P＜0.05); 100 μmol/L MPEP处理U251细胞4、8、12、24 h后磷酸化c-Jun、磷酸化JNK表达量均降低,而且随着用药时间的延长,U251细胞磷酸化c-Jun、磷酸化JNK的表达越低,差异有统计学意义(P＜0.05).结论 mGluR5特异性拮抗剂MPEP能抑制人胶质瘤细胞U251增殖,其机制可能与JNK通路中磷酸化c-Jun及磷酸化JNK的表达水平改变有关,并且MPEP引起的这种改变表现出浓度及时间依赖性.
目的 通過觀察代謝型穀氨痠受體5亞型(mGluR5)的特異性拮抗劑2-甲基-6-苯基乙炔嘧啶(MPEP)對人膠質瘤細胞U251的影響間接探討mGluR5對U251增殖的影響併對其機製進行分析.方法 體外常規培養人膠質瘤細胞株U251,噻唑藍(MTT)檢測100 μmol/L MPEP、20μmol/LSP600125(JNK特異性拮抗劑)、400感染複數(MOI) Ad-△ 169(c-Jun負顯性病毒)作用48 h後U251細胞增殖的變化;Western blotting檢測5、10、20、50、100 μmol/LMPEP作用24 h及100μmol/L MPEP作用4、8、12、24 h後U251細胞錶達燐痠化c-Jun及燐痠化JNK的變化.結果 1 00 μmol/L MPEP、20 μmol/LSP600125、400MOIAd-△169處理48 h後U251細胞的吸光度值均明顯低于對照組,差異有統計學意義(P＜0.05);5、10、20、50、100 μmol/L MPEP處理人膠質瘤細胞U251 24 h後的燐痠化c-Jun、燐痠化JNK錶達量均明顯低于對照組,而且隨著用藥濃度的增加,U251細胞燐痠化c-Jun、燐痠化JNK的錶達越低,差異有統計學意義(P＜0.05); 100 μmol/L MPEP處理U251細胞4、8、12、24 h後燐痠化c-Jun、燐痠化JNK錶達量均降低,而且隨著用藥時間的延長,U251細胞燐痠化c-Jun、燐痠化JNK的錶達越低,差異有統計學意義(P＜0.05).結論 mGluR5特異性拮抗劑MPEP能抑製人膠質瘤細胞U251增殖,其機製可能與JNK通路中燐痠化c-Jun及燐痠化JNK的錶達水平改變有關,併且MPEP引起的這種改變錶現齣濃度及時間依賴性.
목적 통과관찰대사형곡안산수체5아형(mGluR5)적특이성길항제2-갑기-6-분기을결밀정(MPEP)대인효질류세포U251적영향간접탐토mGluR5대U251증식적영향병대기궤제진행분석.방법 체외상규배양인효질류세포주U251,새서람(MTT)검측100 μmol/L MPEP、20μmol/LSP600125(JNK특이성길항제)、400감염복수(MOI) Ad-△ 169(c-Jun부현성병독)작용48 h후U251세포증식적변화;Western blotting검측5、10、20、50、100 μmol/LMPEP작용24 h급100μmol/L MPEP작용4、8、12、24 h후U251세포표체린산화c-Jun급린산화JNK적변화.결과 1 00 μmol/L MPEP、20 μmol/LSP600125、400MOIAd-△169처리48 h후U251세포적흡광도치균명현저우대조조,차이유통계학의의(P＜0.05);5、10、20、50、100 μmol/L MPEP처리인효질류세포U251 24 h후적린산화c-Jun、린산화JNK표체량균명현저우대조조,이차수착용약농도적증가,U251세포린산화c-Jun、린산화JNK적표체월저,차이유통계학의의(P＜0.05); 100 μmol/L MPEP처리U251세포4、8、12、24 h후린산화c-Jun、린산화JNK표체량균강저,이차수착용약시간적연장,U251세포린산화c-Jun、린산화JNK적표체월저,차이유통계학의의(P＜0.05).결론 mGluR5특이성길항제MPEP능억제인효질류세포U251증식,기궤제가능여JNK통로중린산화c-Jun급린산화JNK적표체수평개변유관,병차MPEP인기적저충개변표현출농도급시간의뢰성.
Objective To discuss the effects of metabotropic glutamate receptor 5 subtype (mGluR5) on proliferation of U251 cells and the underlying mechanism by observing the effect of antagonist 2-methyl-6-(phenylethynyl)-pyridine (MPEP) on proliferation of U251 cells.Methods Human glioma cell line U251 was conventionally cultured in vitro; MTT assay was employed to detect the effects of 100 μmol/L MPEP,20 μmol/L SP600125 (antagonist to JNK) and 400 multiplicity of infection (MOI) adenoviruses-△169 (negatively dominant virus ofc-Jun) for 48 h on U251 cells.Western blotting was used to observe the expressions of phosphorylated c-Jun and phosphorylated JNK in U251 cells after being treated with different concentrations of MPEP (5,10,20,50 and 100 μmol/L) for 24 h andwith 100 μmol/LMPEP for4,8,12and24h.Results MTT assay showed that U251 cellsbeing treated with 100 μmol/L MPEP,20 μmoUL SP600125 and 400MOI Ad-△169 for 48 h had significantly lower absorbance value than their control groups (P＜0.05).Western blotting indicated that the expressions of phosphorylated c-Jun and phosphorylated JNK in U251 cells being treated with 5,10,20,50 and 100 μmol/L MPEP for 24 h were significantly lower than those in normal controls (P＜0.05); the higher the MPEP concentration,the lower the expression ofphosphorylated c-Jun.The expressions of phosphorylated c-Jun and phosphorylated JNK in U251 cells being treated with 100 μmol/L MPEP for 4,8,12 and 24 h were significantly lower than those in normal controls (P＜0.05); the longer the MPEP given time,the lower the expression ofphosphorylated c-Jun.Conclusions MPEP,the antagonist mGluR5,could significantly promote cell proliferation of U251,which might be relevant to the expressions of phosphorylated c-Jun and phosphorylated JNK in JNK pathway,and the trend is concentration-dependent and time-dependent.
