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目的应用比较蛋白质组学方法分析小细胞肺癌(SCLC)与其配对的正常肺组织差异表达蛋白质,为阐明SCLC发病机制、筛选其早期诊断标志物提供有益的思路。方法应用双相电泳(2-DE)分离6例SCLC及其配对的正常肺组织可溶性总蛋白;应用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)获得差异蛋白点的肽质量指纹图谱(PMF);通过Mascot软件查询NCBI或SWISS-PROT数据库鉴定蛋白质。结果每张图谱约检测到800多个蛋白质点。经匹配分析,肺癌组织之间和正常肺组织之间凝胶图像的匹配率分别为75.5%和78.2%;选择14个背景清晰、重复性及分辨率较好、蛋白表达差异明显的点进行质谱分析,结合表观分子量及等电点(pI),初步鉴定了11种(六类)蛋白质:①与蛋白质降解通路有关的蛋白质:蛋白酶体α亚单位3型、蛋白酶体β亚单位2型及β亚单位5型;②自由基/抗氧化剂类:锰-超氧化物歧化酶(Mn-SOD)、过氧化氢酶(CAT)和黄素还原酶(FR);③细胞骨架类:原肌球蛋白-3(Tpm-3);④与能量代谢有关的蛋白质:硫氧还蛋白过氧化物酶(TPX1);⑤分子伴侣:抑制素(PHB),内质网蛋白ER29(Erp29);⑥其他:可溶性NSF黏附蛋白(alpha SNAP)。结论应用2-DE及MALDI-TOF-MS方法分离并初步鉴定了11种(六类)蛋白质;这些蛋白质与SCLC发生发展密切相关,部分可能成为SCLC诊断及治疗的分子靶点。
Objective To analyze the differentially expressed proteins of small cell lung cancer (SCLC) paired with normal lung tissue using comparative proteomics methods, which will provide useful ideas for elucidating the pathogenesis of SCLC and screening its early diagnostic markers. Methods Two-phase electrophoresis (2-DE) was used to isolate soluble total proteins in 6 normal SCLC and its matched normal lung tissues. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS) was used to obtain peptide mass fingerprints of differential protein spots. (PMF); Identification of proteins by querying the NCBI or SWISS-PROT database using Mascot software. As a result, more than 800 protein spots were detected per map. After matching analysis, the matching rates of the gel images between lung cancer tissues and normal lung tissues were 75.5% and 78.2%, respectively; 14 points with clear background, good repeatability and resolution, and significant differences in protein expression were selected for mass spectrometry. Analysis, combined with apparent molecular weight and isoelectric point (pI), initially identified 11 (six) proteins: 1 Proteins associated with protein degradation pathways: proteasome subunit 3, proteasome subunit 2 and β subunit 5 type; 2 free radicals/antioxidants: Manganese-superoxide dismutase (Mn-SOD), catalase (CAT) and flavin reductase (FR); 3 Cytoskeleton class: Primary muscle Globulin-3 (Tpm-3); 4 proteins involved in energy metabolism: thioredoxin peroxidase (TPX1); 5 chaperones: inhibin (PHB), endoplasmic reticulum protein ER29 (Erp29); 6 Others: Soluble NSF adhesion protein (alpha SNAP). Conclusion The 2-DE and MALDI-TOF-MS methods were used to isolate and preliminarily identify eleven (six) proteins; these proteins are closely related to the occurrence and development of SCLC and may become molecular targets for the diagnosis and treatment of SCLC.