人参皂苷Rg3对肺癌细胞蛋白表达的影响研究  

Effect of 20(R) ginsenoside Rg3 on protein expression of lung cancer cell line

作　　者：安宁[1] 朱文[1] 冯志华[1] 叶苏娟[1] 余川江[1] 蔡春季[1] 

机构地区：[1]四川大学华西医院生物治疗国家重点实验室,成都610041

出　　处：《中国肺癌杂志》2008年第3期311-320,共10页Chinese Journal of Lung Cancer

基　　金：四川大学华西医院留学回国人员基金资助(No.2005)~~

摘　　要：背景与目的肺癌是对人类健康和生命危害最大的恶性肿瘤。肺癌转移是肺癌的恶性标志和特征,也是肺癌病人治疗失败和死亡的主要原因。人参皂苷Rg3具有明显的抗肿瘤效应,该研究将应用双向凝胶电泳技术探讨人参皂苷Rg3抗肺癌侵袭转移分子作用机制。方法筛选人参皂苷Rg3作用于人肺巨细胞癌高转移细胞株的半抑制浓度(IC50),用0.1倍半抑制浓度(0.1×IC50)的Rg3溶液作用人肺巨细胞癌高转移细胞株95D72h,利用固相PH梯度双向凝胶电泳分离Rg3处理前后人肺巨细胞癌高转移细胞株95D的总蛋白,用图像分析软件比较分析Rg3处理前后细胞间的差异表达蛋白质。对15个差异表达的蛋白质点通过MALD-TOF-MS/MS,LC-MS/MS和蛋白质数据库检索进行鉴定。结果人参皂苷Rg3作用人肺巨细胞癌高转移细胞株95D的半抑制浓度为100μg/mL。有12个蛋白点仅在对照组细胞株中出现,有6个蛋白点仅在药物处理组出现;在对照组与药物处理组中都存在,但在对照组中高表达的蛋白点有7个,在药物处理组中高表达的蛋白点有2个。通过对差异明显的15个蛋白质点进行质谱鉴定和蛋白质数据库查询,发现氯离子通道蛋白1(Chloride intracellular channel protein1)、泛素结合酶E2-25Kda(Ubiquitin-Conjugating Enzyme E2-25Kda)等蛋白只在对照组有表达;14-3-3θ蛋白(14-3-3protein theta)、Ski作用蛋白(SKI-interacting protein)只在药物处理组有表达;膜联蛋白A2(annexin A2)、肌动蛋白结合蛋白Ⅱ同构体b(profilin 2 isoformb)等蛋白在对照组的表达量显著高于药物处理组;14-3-3ζ蛋白(14-3-3 protein zeta),真核翻译起始因子4H(Eukaryotic translation initiation factor 4H)在药物处理组的表达量显著高于对照组。结论该研究结果提示Rg3处理前后人肺巨细胞癌高转移细胞株蛋白质组的表达存在明显差异,鉴定的差异蛋白质多与肿瘤的侵袭转移相关。这些差异蛋白质有可能为�Background and objective Lung cancer is the most dangerous threating tumor to human＇s health and life. Metastasis is not only the malignant characteristics of lung cancer, hut also the chief cause of failure to cure the disease and of high mortality. Ginsenoside Rg3 has been proved to have obvious effect against tumor. The molecular mechanism of Rg3 against lung cancer cell line will be investigated by using two-dimensional gel electrophoresis in this paper. Methods The IC50 of Rg3 against human high-metastatic large cell lung cancer cell line 95D was determined by MTT. Then 95D cells were treated with Rg3 at the concentration of 0.1 ×IC50 for 72 h. The total proteins of 95D cell line treated with Rg3 and not treated were separated and protein profiles were obtained by using immobilized PH gradient （IPG）-based two-dimensional gel electrophoresis. The differential expression proteins of 95D cell line treated with Rg3 and not treated were analyzed using image analysis software. 15 of differentially expressed proteins were further identified using MALDI-TOF MS/MS analysis and LC-MS/MS analysis. Protein identification was performed by searching the protein database. Results The IC50 of Rg3 against 95D cell line was 100 μg/mL. There were 27 differently expressed protein spots through analysis by Image Master Microsoft. 15 proteins were identified using mass spectrometry. Chloride intracellular channel protein 1, Ubiquitin-Conjugating Enzyme E2-25 Kda only expressed in control; 14-3-3 protein teta, SKI-interacting protein only expressed in 95D cell line treated by Rg3; Annexin A2, profilin 2 isoform b were downregulated in 95D cell line treated by Rg3; 14-3-3 protein zeta, Eukaryotic translation initiation factor 4H were upregulated in 95D cell line treated by Rg3. Conclusion A significantly different expression of proteins were found in 95D cell line treated with Rg3 and those not treated. Most of identified proteins have been reported to be associated with tumor metastasis. The identified proteins will provid

关 键 词：人参皂苷RG3 蛋白质表达 双向凝胶电泳 质谱 

分 类 号：R734.2[医药卫生—肿瘤]