CAJ | 학술논문

目的：探讨苦参碱对人横纹肌肉瘤（RMS）RD细胞ERK通路的影响。方法取对数生长期的细胞，随机分为对照组（含细胞无药物）及3个药物干预组（分别为U012610μmol组、苦参碱1.0 mg/mL组和U012610μmol与苦参碱1.0 mg/mL共同作用组）。蛋白质印迹法（Western blotting）检测p44/p42MAPK（ERK1/2）及Phospho_p44/p42MAPK （p_ERK1/2）的表达；反转录聚合酶链反应（RT_PCR）方法检测erk mRNA的表达。最后采用SPSS 16.0统计软件进行单因素方差分析。结果 Western blotting法检测ERK1/2、p_ERK1/2的表达，对照组（U012610μmol组，苦参碱1.0 mg/mL组及U012610μmol与苦参碱1.0 mg/mL共同作用组）的灰度值分别为：2.84±0.03、2.64±0.03、2.14±0.01、2.10±0.02及1.76±0.15、1.37±0.19、1.21±0.24、1.19±0.22。与对照组比较，苦参碱1.0 mg/mL组及苦参碱与U0126共同作用组对ERK1/2、p_ERK1/2蛋白表达量明显降低，差异均有统计学意义（P<0.01）；RT_PCR方法检测erk mRNA的表达，对照组（U012610μmol组，苦参碱1.0 mg/mL组及U012610μmol与苦参碱1.0 mg/mL共同作用组）的灰度比值分别为0.59±0.04、0.57±0.04、0.25±0.02和0.18±0.01，苦参碱1.0 mg/mL组及苦参碱与U0126共同作用组erk mRNA的表达量较对照组明显降低，差异均有统计学意义（P<0.01）。结论苦参碱在体外环境下作用RMS_RD细胞，可抑制erk mRNA的表达及ERK1/2蛋白磷酸化的活性。
목적：탐토고삼감대인횡문기육류（RMS）RD세포ERK통로적영향。방법취대수생장기적세포，수궤분위대조조（함세포무약물）급3개약물간예조（분별위U012610μmol조、고삼감1.0 mg/mL조화U012610μmol여고삼감1.0 mg/mL공동작용조）。단백질인적법（Western blotting）검측p44/p42MAPK（ERK1/2）급Phospho_p44/p42MAPK （p_ERK1/2）적표체；반전록취합매련반응（RT_PCR）방법검측erk mRNA적표체。최후채용SPSS 16.0통계연건진행단인소방차분석。결과 Western blotting법검측ERK1/2、p_ERK1/2적표체，대조조（U012610μmol조，고삼감1.0 mg/mL조급U012610μmol여고삼감1.0 mg/mL공동작용조）적회도치분별위：2.84±0.03、2.64±0.03、2.14±0.01、2.10±0.02급1.76±0.15、1.37±0.19、1.21±0.24、1.19±0.22。여대조조비교，고삼감1.0 mg/mL조급고삼감여U0126공동작용조대ERK1/2、p_ERK1/2단백표체량명현강저，차이균유통계학의의（P<0.01）；RT_PCR방법검측erk mRNA적표체，대조조（U012610μmol조，고삼감1.0 mg/mL조급U012610μmol여고삼감1.0 mg/mL공동작용조）적회도비치분별위0.59±0.04、0.57±0.04、0.25±0.02화0.18±0.01，고삼감1.0 mg/mL조급고삼감여U0126공동작용조erk mRNA적표체량교대조조명현강저，차이균유통계학의의（P<0.01）。결론고삼감재체외배경하작용RMS_RD세포，가억제erk mRNA적표체급ERK1/2단백린산화적활성。
Objective To explore the influence of Matrine on MAPK/ERK signaling pathways of human rhabdomyosarcoma (RMS) RD cells. Methods The cells at logarithm growth phase were chosen and randomly divided into the control group (in_cluding cells without Matrine)and three intervention groups of different concentrations(respectively for U0126 10 um,Matrine 1.0 mg/mL,U0126 10 um combined with Matrine 1.0 mg/mL). The expressions of p44/p42MAPK (ERK1/2) and Phosopho_p44/p42MARK(p_ERK1/2) were detected by Western blotting,and the expression of erk mRNA was detected by reverse transcription_polymerase chain reaction(RT_PCR). One_way analysis of variance was performed by SPSS16.0 statistical software. Results The grey values of expression of ERK1/2,p_ERK1/2 in the control group,U0126 10μmol group,Matrine 1.0 mg/mL group and U0126 10μmol combined with Matrine 1.0 mg/mL group were(2.84±0.03),(2.64±0.03),(2.14±0.01),(2.10±0.02);(1.76±0.15), (1.37±0.19),(1.21±0.24),(1.19±0.22) by Western blotting respecitvely,compared with the control group,the Matrine 1.0 mg/mL and Matrine 1.0 mg/mL combined U0126 10μmol groups ERK1/2,p_ERK1/2 protein expressions were statistically significant difference(P<0.01);The expression of erkmRNA in the control group,U0126 10 μmol group,Matrine 1.0 mg/mL group and U0126 10μmol combined with Matrine 1.0 mg/mL group were(0.59±0.04),(0.57±0.04),(0.25±0.02)and(0.18±0.01) by RT_PCR, significant differences were found in the Matrine 1.0 mg/mL group and expression amount of Matrine 1.0 mg/mL combined U0126 10μmol compared with the control group(P<0.01). Conclusion Matrine may inhibit the RMS_RD cells expression and the phos_phorylated activity of ERKmRNA ERK1/2 protein in vitro.