摘要
背景与目的 肺癌转移是肺癌的恶性标志和特征,也是肺癌患者治疗失败和死亡的主要原因。 为了更好地了解和阐明肺癌转移的分子机制,发现早期诊断和逆转肺癌转移的分子标志物,本研究应用双向 凝胶电泳技术,比较人高转移大细胞肺癌细胞株(L9981)和人低转移大细胞肺癌细胞株(NL9980)间蛋白质组 表达的差异。方法 利用固相pH梯度双向凝胶电泳,分离L9981和NL9980细胞株的总蛋白,用图像分析 软件比较分析并识别细胞间的差异表达蛋白质。结果 应用比较蛋白质组学技术对L9981和NL9980细胞 株的蛋白质表达进行双向电泳分离,成功地获得了蛋白组分辨率高、重复性好的双向凝胶电泳图谱。在三次 重复实验结果中,在L9981和NL9980细胞株中检测到的蛋白质点数分别为902±169和941±173,蛋白质斑 点位置在IEF方向的平均偏差为(0.858±0.076)mm,在SDS PAGE方向的平均偏差为(1.514±0.127)mm, 蛋白质表达量的变异为12.06%～12.22%。经ImageMaster软件分析后发现有15个蛋白质点仅在L9981 肺癌细胞株中检测到有表达,27个蛋白质点仅在NL9980中检测到有表达。发现4个蛋白质点在两种肺癌 细胞株中均存在,但在两种细胞株间的表达量差异在2倍以上。
Background and objective Metastasis is not only the malignant characteristics of lung can- cer, but also the chief cause of failure to cure and high mortality of lung cancer. To better explore and understand the mechanism of lung cancer metastasis and to search for potential markers for early diagnosing and reversing lung cancer metastasis, differential proteomic analysis is conducted in two human large cell lung cancer cell lines with high metastasis potentials (L9981) and low metastasis potentials (NL9980) by two-dimension gel electrophoresis (2-DE). Methods The total proteins of the two cell lines were separated by immobilized pH gradient (IPG)-based 2-DE. The differentially expressed proteins of the two cell lines were analyzed using image analysis software. Results A high resolution and reproducible 2-DE image was successfully obtained. Average deviations for protein position in IEF direction were (0.858±0.076)mm and (1.514±0.127)mm in SDS-PAGE direction. The relative standard deviation for protein volume was (12.06±0.580)% in L9981 and (12.22±0.640)% in NL9980. The average total number of protein spots was 902±169 in L9981 cells and 941±173 in NL9980 cells in three repeated experiments. Image analysis of siliver-stained 2-DE image revealed that 4 protein spots had significant differential expressions in L9981 and NL9980 (student’s t-test, P<0.05). Fifteen protein spots were only detected in L9981, and 27 protein spots were only detected in NL9980. Conclusion The results in this study suggest that an obviously differential proteomic expression exists between the human high- and low-metastatic large cell lung cancer cell lines. It will be helpful to further understand the molecular mechanisms of lung cancer invasion and metastasis, and provide new experimental evidence for searching metastatic-related molecule of lung cancer.
出处
《中国肺癌杂志》
CAS
2005年第1期1-7,共7页
Chinese Journal of Lung Cancer
基金
国家自然科学基金重点项目(No.30430300)
国家自然科学基金(No.30070333)
教育部留学回国人员基金([2004]527)
四川大学青年科学基金(No.200349)资助~~
关键词
蛋白质组
双向凝胶电泳
转移
人大细胞肺癌细胞株
差异表达
Proteome Two-dimension gel electrophoresis Metastasis Human large cell lung cancer cell line Differential expression
