Cancer Res Treat. 2003 Dec;35(6):489-96. doi: 10.4143/crt.2003.35.6.489.
Small cell lung cancer is one of the major causes of death from cancer worldwide. To explore the expressions of global protein in small cell lung cancer cells, a proteomic approach, to identify the proteins, was used and the establishment of a protein reference map attempted.
Two-dimensional gel electrophoresis (2-DE), with subsequent analysis by mass spectrometry (MS), was applied to the study of protein identification from a small cell lung cancer cell line, NCI- H211. The cells were lysed, and the extracts subjected to isoelectric focusing, with immobilized pH gradients, followed by second dimension SDS-PAGE. The polypeptides were identified by peptide mass fingerprinting, with MALDI-TOF MS, after in-gel protein digestion.
From silver staining of the gel, around two thousands protein spots were separated by the 2-DE. Of these protein spots visualized in the gel, one hundred and ten were identified by peptide mass fingerprinting. Different proteins, such as enzymes, cytoskeletal proteins and proteins common to eukaryotic cells, were identified.
The protein expressions of the small cell lung cancer cells were analyzed to establish a protein reference map. The reference map presented here may serve as a working tool for the further study of small cell lung cancer.
小细胞肺癌是全球癌症死亡的主要原因之一。为了探索小细胞肺癌细胞中的整体蛋白质表达情况,采用了蛋白质组学方法来鉴定蛋白质,并试图建立蛋白质参考图谱。
应用二维凝胶电泳(2-DE),随后通过质谱(MS)分析,从小细胞肺癌细胞系 NCI-H211 中研究蛋白质鉴定。将细胞裂解,提取物进行等电聚焦,使用固定 pH 梯度,然后进行 SDS-PAGE 第二维电泳。通过胶内蛋白消化,MALDI-TOF MS 进行肽质量指纹图谱分析,鉴定多肽。
通过凝胶银染,通过 2-DE 分离出约两千个蛋白质斑点。在凝胶中可见的这些蛋白质斑点中,有 110 个通过肽质量指纹图谱鉴定。鉴定出了不同的蛋白质,如酶、细胞骨架蛋白和真核细胞共有的蛋白质。
分析了小细胞肺癌细胞的蛋白质表达情况,建立了蛋白质参考图谱。本文提供的参考图谱可作为进一步研究小细胞肺癌的工作工具。