摘要
目的:探讨定量PCR和Western blot结合检测对Lewis肺癌C57小鼠瘤细胞表皮生长因子受体(EGFR)、NF-κB表达影响,观察该方对调节癌细胞信号转导作用机制。方法:小鼠自接种瘤株细胞24 h后称重、编号,随机分为8组,每组5只,分别为:1回药高剂量组(AKH);2荷瘤模型组(对照组);3回药中剂量组(AKM);4单纯化疗药组(CTX组);5回药中剂量组加化疗组(AKM+CTX);6回药低剂量加化疗药组(AKL+CTX);7回药低剂量组(AKL);8回药高剂量加化疗药组(AKH+CTX)。采用RT-PCR和western blot方法检测小鼠组织样本中EGFR、NF-κBm RNA转录水平和蛋白质表达的变化情况。结果:实时荧光定量PCR结果显示:与模型组小鼠比较,给予回药爱康方,高、中、低剂量干预后,小鼠瘤细胞中EGFR m RNA表达水平显著下调(P<0.01),而低剂量对NF-κBm RNA的表达水平无统计学差异(P>0.05),给予回药爱康方,高+CTX、中+CTX、低+CTX、CTX后,小鼠瘤细胞中EGFR表达水平显著下调(P<0.01),小鼠瘤细胞中NF-κBm RNA表达水平显著上调(P<0.01);与模型组比较,给予回药爱康方高、中、低剂量干预后,Lewis肺癌C57小鼠EGFR、NF-κB蛋白表达水平无统计学差异性(P>0.05),给予回药爱康方高+CTX、中+CTX、低+CTX、CTX干预后,Lewis肺癌C57小鼠EGFR蛋白表达水平显著下调、NF-κB蛋白表达水平显著上调(P<0.01或P<0.05)。结论:回药爱康方高、中、低与回药开康方高+CTX、中+CTX、低+CTX、CTX均对EGFRm RNA的表达水平有影响,仅低剂量对NF-κBm RNA的表达水平无影响,同时回药爱康方高、中、低剂量均对EGFR、NF-κB蛋白表达无影响。
Objective： To explore the effect of Hui AKF on lung cancer tumor cells EGFR and NF-κb expression by quantitative PCR and Western blot and observe the mechanism of regulating cancer cell signal transduction.Methods：Mice were inoculated tumor cell strains within 24 h after weighing and serially numbering,and were randomly divided into8 groups,5 in each group,respectively：（1） AKF high dose group（AKH）;（2） a tumor-burdened model group（control group）;（3） AKF medium dose group（AKM）;（4） the simple effective group（CTX）;（5） AKF low dose and chemotherapy group（AKM ＋ CTX）;（6） AKF low dose and effective group（AKL ＋ CTX）;（7） AKF low dose group（AKL）;（8）AKF high dose and effective group（AKH ＋ CTX）,by using RT-PCR and Western blot method to detect tissue samples of EGFR in mice,the NF-kappa B m RNA transcription level and protein expression changes.Results：The real-time fluorescent quantitative PCR results showed compared with model group＇s mice,after AKF high,medium and low doses intervention,EGFR m RNA expressions in tumor cells of mice were significantly raised（P〈0.01） while low dose on the NF-kappa B m RNA expression level had no statistical difference（P〈0.05）.And after intervention respectively by AKH ＋CTX,AKM ＋ CTX and AKL ＋ CTX and CTX,EGFR in tumor cells in mice was decreased and the NF-kappa B m RNAexpression level increased significantly（P〈0.01）.Compared with model group,after intervention by high,medium and low dose AKF,Lewis lung EGFR C57 mice NF-kappaB protein expression level had no statistical difference（P〈0.05）.After intervention by respectively by AKH ＋ CTX,AKM ＋ CTX and AKL ＋ CTX and CTX,Lewis lung EGFR C57 mice NF-kappaB protein expression levels increased significantly（P〈0.01 or P〈0.05）.Conclusion： AKF high,medium and low doses and the three levels of doses respectively combined with CTX all influence the expression level of EGFRmR NA.Only low dose of AKF has on effect on NF-kappaB mR NA expression level and at the same time the three levels of doses have no effect on expressions of EGFR and NF-kappa B protein.
出处
《中华中医药学刊》
CAS
北大核心
2016年第12期2831-2835,I0018,共6页
Chinese Archives of Traditional Chinese Medicine
基金
国家自然科学基金项目(81160462)
