中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2013年
8期
1580-1582
,共3页
穆林%郭芽%李建钦%向旭东%刘绍坤
穆林%郭芽%李建欽%嚮旭東%劉紹坤
목림%곽아%리건흠%향욱동%류소곤
氧化苦参碱%非小细胞肺癌%脱噬作用%c-Jun氨基端激酶通路
氧化苦參堿%非小細胞肺癌%脫噬作用%c-Jun氨基耑激酶通路
양화고삼감%비소세포폐암%탈서작용%c-Jun안기단격매통로
Oxymatrine%Non-small cell lung carcinoma%Apoptosis%Phosphorylated c-Jun N-terminal kinase
目的 观察氧化苦参碱(OXY)对人非小细胞肺癌A549细胞凋亡的影响并探讨其机制.方法 噻唑蓝(MTT)法测定OXY对A549人肺癌细胞活力的作用.流式细胞术测定A549细胞凋亡率.荧光定量聚合酶链反应(FQ-PCR)和Westem blot方法测定OXY对A549细胞B淋巴细胞/白血病-2(bcl-2)、bcl-2相关X蛋白(bax) mRNA和蛋白表达的影响.Western blot方法测定OXY对A549细胞的细胞外信号调节蛋白激酶(ERK)、c-Jun氨基端激酶(JNK)、p38丝裂原活化激酶(p38MAPK)、磷酸化ERK(p-ERK)、p-JNK、p-p38 MAPK蛋白表达的影响.结果 OXY可抑制A549细胞增殖.与对照组[(5.63±0.76)％]比较,OXY组细胞凋亡率[(13.26±2.15)％]上升(P＜0.01)；与对照组(1.02±0.27、0.23±0.04)比较,OXY组bax mRNA(1.61±0.19)和蛋白(0.71±0.10)表达上升(P＜0.05,P＜0.01)；与对照组比较(0.94 ±0.14、0.64±0.09),OXY组bcl-2mRNA(0.42±0.05)和蛋白(0.21±0.04)表达下降(P＜0.01).与对照组(0.29±0.03)比较,OXY组p-JNK表达水平(0.66 ±0.06)明显升高(P＜0.01),其他蛋白表达均无明显变化.与OXY组(0.66±0.07、0.28±0.03、0.65±0.08)比较,同时给予OXY和p-JNK抑制剂SP600125组bax蛋白表达(0.39±0.05)下降,bcl-2表达(0.50±0.08)上升,p-JNK蛋白(0.26 ±0.04)表达下降(P＜0.01).结论 OXY碱可诱导A549细胞凋亡,可能与升高p-JNK水平有关.
目的 觀察氧化苦參堿(OXY)對人非小細胞肺癌A549細胞凋亡的影響併探討其機製.方法 噻唑藍(MTT)法測定OXY對A549人肺癌細胞活力的作用.流式細胞術測定A549細胞凋亡率.熒光定量聚閤酶鏈反應(FQ-PCR)和Westem blot方法測定OXY對A549細胞B淋巴細胞/白血病-2(bcl-2)、bcl-2相關X蛋白(bax) mRNA和蛋白錶達的影響.Western blot方法測定OXY對A549細胞的細胞外信號調節蛋白激酶(ERK)、c-Jun氨基耑激酶(JNK)、p38絲裂原活化激酶(p38MAPK)、燐痠化ERK(p-ERK)、p-JNK、p-p38 MAPK蛋白錶達的影響.結果 OXY可抑製A549細胞增殖.與對照組[(5.63±0.76)％]比較,OXY組細胞凋亡率[(13.26±2.15)％]上升(P＜0.01)；與對照組(1.02±0.27、0.23±0.04)比較,OXY組bax mRNA(1.61±0.19)和蛋白(0.71±0.10)錶達上升(P＜0.05,P＜0.01)；與對照組比較(0.94 ±0.14、0.64±0.09),OXY組bcl-2mRNA(0.42±0.05)和蛋白(0.21±0.04)錶達下降(P＜0.01).與對照組(0.29±0.03)比較,OXY組p-JNK錶達水平(0.66 ±0.06)明顯升高(P＜0.01),其他蛋白錶達均無明顯變化.與OXY組(0.66±0.07、0.28±0.03、0.65±0.08)比較,同時給予OXY和p-JNK抑製劑SP600125組bax蛋白錶達(0.39±0.05)下降,bcl-2錶達(0.50±0.08)上升,p-JNK蛋白(0.26 ±0.04)錶達下降(P＜0.01).結論 OXY堿可誘導A549細胞凋亡,可能與升高p-JNK水平有關.
목적 관찰양화고삼감(OXY)대인비소세포폐암A549세포조망적영향병탐토기궤제.방법 새서람(MTT)법측정OXY대A549인폐암세포활력적작용.류식세포술측정A549세포조망솔.형광정량취합매련반응(FQ-PCR)화Westem blot방법측정OXY대A549세포B림파세포/백혈병-2(bcl-2)、bcl-2상관X단백(bax) mRNA화단백표체적영향.Western blot방법측정OXY대A549세포적세포외신호조절단백격매(ERK)、c-Jun안기단격매(JNK)、p38사렬원활화격매(p38MAPK)、린산화ERK(p-ERK)、p-JNK、p-p38 MAPK단백표체적영향.결과 OXY가억제A549세포증식.여대조조[(5.63±0.76)％]비교,OXY조세포조망솔[(13.26±2.15)％]상승(P＜0.01)；여대조조(1.02±0.27、0.23±0.04)비교,OXY조bax mRNA(1.61±0.19)화단백(0.71±0.10)표체상승(P＜0.05,P＜0.01)；여대조조비교(0.94 ±0.14、0.64±0.09),OXY조bcl-2mRNA(0.42±0.05)화단백(0.21±0.04)표체하강(P＜0.01).여대조조(0.29±0.03)비교,OXY조p-JNK표체수평(0.66 ±0.06)명현승고(P＜0.01),기타단백표체균무명현변화.여OXY조(0.66±0.07、0.28±0.03、0.65±0.08)비교,동시급여OXY화p-JNK억제제SP600125조bax단백표체(0.39±0.05)하강,bcl-2표체(0.50±0.08)상승,p-JNK단백(0.26 ±0.04)표체하강(P＜0.01).결론 OXY감가유도A549세포조망,가능여승고p-JNK수평유관.
Objective To investigate the effects of oxymatrine (OXY) on apoptosis and the regulatory mechanism in the human non-small cell lung carcinoma A549 cells.Methods A549 cells were cultured.Methyl thiazol tetrazolium (MTT) assay was used to determine cell proliferation.The apoptosis rate was detected by using flow cytometry.Real-time fluorescence quantitative polymerase chain reaction (FQ-PCR) and Western blotting were used to examine OXY mRNA and protein expression.Extracellular signal-regulated kinase (ERK),C-Jun N-terminal kinase (JNK),p38 mitogen-activated protein kinase (p38MAPK),p-ERK,p-JNK and p-p38MAPK protein expression levels were tested by using Western blotting.Results OXY could inhibit the proliferation of A549 cells.As compared with control group [(5.63 ±0.76)％],the apoptosis rate was increased [(13.26 ± 2.15)％] in OXY group (P ＜ 0.01).As compared with control group (1.02 ±0.27,0.23 ±0.04),B lymphocytes/leukemia-2 (bcl-2) associated X protein (bax) mRNA (1.61 ±0.19) and protein (0.71 ±0.10) expression levels were increased in OXY group (P ＜ 0.05,P ＜ 0.01).As compared with control group (0.94 ± 0.14,0.64 ± 0.09),bcl-2 mRNA (0.42 ± 0.05) and protein (0.21 ± 0.04) expression levels were decreased in OXY group (P ＜0.01).As compared with control group (0.29 ± 0.03),the expression of p-JNK was significantly increased in OXY group (0.66 ±0.06) (P ＜0.01).bax expression (0.39 ±0.05) and p-JNK (0.26 ±0.04) were reduced,and bcl-2 expression (0.50 ± 0.08) was increased in OXY + SP600125 group as compared with those in OXY group (0.66 ± 0.07,0.28 ± 0.03 and 0.65 ± 0.08 respectively) (P ＜0.01).Conclusion OXY could induce apoptosis of A549 cells by up-regulating p-JNK pathway.
