人肺鳞癌组织及癌旁组织蛋白质双向电泳图谱的差异分析

背景与目的:肺鳞癌的发生发展是一个多因素、多步骤的过程,虽然从基因和转录水平已对肺癌进行了许多成功的研究,但其癌变机制仍不十分明确,目前尚缺乏有效的用于肺鳞癌早期诊断和预后监测的特异性分子标志物。本研究的目的是利用蛋白质组学方法,建立分辨率高和重复性好的人肺鳞癌组织及其癌旁正常支气管上皮组织的双向凝胶电泳图谱,并识别鉴定其差异表达的蛋白质。方法:利用固相pH梯度(immobilizedpHgradient,IPG)双向凝胶电泳(two-dimensionalelectrophoresis,2-DE),分离人肺鳞癌组织及癌旁正常支气管上皮组织的总蛋白质,用图像分析软件比较分析,以识别差异表达的蛋白质;应用质谱仪(massspectrometry)得到相应的肽质指纹图谱(peptidemassfingerprint,PMF),然后搜索数据库鉴定部分差异蛋白质点。结果:(1)建立双向凝胶电泳图谱:癌组织和癌旁正常支气管上皮组织3块凝胶的平均蛋白质点数分别为1349±67和1297±73,平均匹配的点数分别为1235±48和1183±56,匹配率达91.5%和91.2%;同一癌组织的3块胶在蛋白质点位置上有较好的重复性,不同胶间蛋白质点在等电聚焦(Isoelectricfocusing,IEF)方向的偏差是(0.873±0.125)mm,在十二烷基磺酸钠聚丙烯酰胺凝胶电泳SDS-PAGE方向上的偏差为(1.025±0.213)mm。(2)肽质指     

Comparative Proteome Analysis of Human Lung Squamous Carcinoma Tissue

目的利用蛋白质组学方法建立人肺鳞癌组织及其癌旁正常支气管上皮组织的差异蛋白质表达谱。方法对20例人肺鳞癌组织和配对的癌旁正常支气管上皮组织进行比较蛋白质组学研究,即利用双向凝胶电泳(2-DE)分离二者总蛋白质后,经图像分析识别差异表达的蛋白,应用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)鉴定差异蛋白质。结果(1)比较分析20例肺鳞癌及正常配对组织的2-DE图谱,找到差异蛋白质点76个;(2)对68个差异蛋白质点进行了肽质指纹图分析,鉴定出一些与瘤基因、细胞周期调控、信号转导等有关的肺鳞癌相关蛋白;(3)肺鳞癌相关蛋白mdm2、c-jun和EGFR存人肺鳞癌组织中高表达,而在正常对照中均表达下调,与蛋白质组的分析鉴定结果是一致的。结论成功鉴定了68个肺鳞癌相关蛋白,为进一步筛选用于肺鳞癌诊断、治疗和预后评估的肺鳞癌分子标志物奠定了坚实的基础。     

蛋白质组学方法筛选早期肺鳞癌相关蛋白

目的:应用蛋白质组学技术筛选早期肺癌癌变相关蛋白.方法:利用双向凝胶电泳方法分离人早期肺鳞癌组织和相应癌旁正常组织总蛋白,选择差异蛋白质点进行质谱分析.免疫组织化学法验证部分差异蛋白质的表达差异.结果:肺鳞癌组织与癌旁正常组织的双向凝胶电泳图谱中平均蛋白质点数分别为626和602个.选择在癌组织中高表达的10个差异蛋白点进行质谱分析,最终鉴定为膜联蛋白1 (annexin-1, Anx-A1)、热休克蛋白27(heat shock protein 27, HSP27)等与细胞周期、信号转导等功能相关的蛋白质.免疫组织化学检测结果显示,Anx-A1和HSP27蛋白在肺鳞癌组织中的阳性表达率均显著增高(P<0.05).结论:蛋白质组学方法是一种应用于初步筛选早期肺癌相关蛋白的有效方法,所鉴定的蛋白为进一步筛选用于肺癌早期诊断及其治疗的分子标志物奠定了前期基础.     

人支气管上皮组织癌变各阶段2-DE图谱及差异分析

支气管上皮细胞的癌变是一个多基因参与、多阶段的复杂过程,但癌变机理仍不清楚,应用蛋白质组学技术研究此过程有可能识别癌变相关蛋白质,对揭示肺鳞癌癌变机制具有重要的意义。本研究的目的是优化支气管上皮组织的蛋白质样品制备方法,建立人支气管上皮癌变各阶段组织的2-DE图谱并进行差异分析,为鉴定肺鳞癌癌变相关蛋白质奠定基础。     

人肺鳞癌组织的比较蛋白质组学研究

目的利用蛋白质组学方法建立人肺鳞癌组织及其癌旁正常支气管上皮组织的差异蛋白质表达谱。方法对20例人肺鳞癌组织和配对的癌旁正常支气管上皮组织进行比较蛋白质组学研究,即利用双向凝胶电泳（2-DE）分离二者总蛋白质后,经图像分析识别差异表达的蛋白,应用基质辅助激光解吸电离飞行时间质谱（MALDI—TOF—MS）鉴定差异蛋白质。结果（1）比较分析20例肺鳞癌及正常配对组织的2-DE图谱,找到差异蛋白质点76个;（2）对68个差异蛋白质点进行了肽质量指纹图分析,鉴定出一些与瘤基因、细胞周期调控、信号转导等有关的肺鳞癌相关蛋白;（3）肺鳞癌相关蛋白mdm2、c-Jun和表皮生长因子受体（EGFR）在人肺鳞癌组织中高表达,而在正常对照中均表达下调,与蛋白质组的分析鉴定结果是一致的。结论成功鉴定了68个肺鳞癌相关蛋白,为进一步筛选用于肺鳞癌诊断、治疗和预后评估的肺鳞癌分子标志物奠定了坚实的基础。     

Membrane proteomic analysis comparing squamous cell lung cancer tissue and tumour-adjacent normal tissue

Lung cancer is the leading cause of cancer-related deaths worldwide. Squamous cell carcinoma is one of the predominant histological subtypes of lung cancer. Detecting lung cancer at an early stage is essential for successful therapy and increasing survival. There are still no satisfactory biomarkers for the early detection of lung cancer. In this study, tumour tissue paired with tumour-adjacent normal bronchial epithelial tissue was obtained from patients with squamous cell lung carcinoma without metastasis. The proteins extracted from the cell membrane were separated by two-dimensional polyacrylamide gel electrophoresis (2-DE) and were analysed with the Image Master two-dimensional platinum software. Twenty-five significantly different protein spots were selected and identified with matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS). A total of 19 proteins were successfully identified. Twelve proteins were up-regulated, and seven proteins were down-regulated in the cancerous tissue compared with the tumour-adjacent normal tissue. One up-regulated protein and one down-regulated protein in squamous cell lung carcinoma were verified by Western blot analysis and RT-PCR; the results were consistent with the 2-DE analysis. In conclusion, membrane proteomics identified a number of candidate biomarker proteins that were differentially expressed between squamous cell lung cancer tissue and adjacent normal tissue. These biomarker candidates have the potential to elucidate the underlying pathogenesis of squamous cell lung cancer.     

双向凝胶电泳飞行时间质谱技术筛选食管癌相关蛋白

目的：建立分辨率高和重复性好的人食管癌组织及其癌旁正常上皮组织的双向凝胶电泳分析方法,并筛选食管癌相关蛋白质。方法：选取10例食管癌及其癌旁正常上皮组织,提取总蛋白质进行双向凝胶电泳分析,并采用Blue silver法染色。凝胶图像分析后,对差异蛋白质进行MALDI-TOF质谱分析。结果：在所鉴定的6个差异蛋白质在中,磷酸丙糖异构酶1、锰超氧化物歧化酶和热休克蛋白27在食管癌组织中表达上调,而鳞状细胞癌抗原1、细胞角蛋白4和膜联蛋白Ⅰ在食管癌中表达下调。结论：所鉴定的这些差异蛋白将为阐明食管癌发生机制提供线索,也为寻找食管癌潜在标志奠定理论基础。     

核基质蛋白LaminA/C在正常乳腺和乳腺癌组织中的差异表达*

目的:采用比较蛋白质组学方法研究正常乳腺和乳腺癌组织中的差异表达核基质蛋白（nuclear matrix protein ,NMP）,并在蛋白质水平对差异蛋白LaminA/C进一步地确证。方法:分别提取4 例正常乳腺和乳腺癌组织的核基质蛋白,进行双向凝胶电泳。分离出的差异表达蛋白质点进行基质辅助激光解吸电离飞行时间质谱（MALDI-TOF-MS）分析,获得肽质量指纹图谱（Peptide Mass Fingerprinting,PMF ）,应用Mascot 搜索引擎在NCBInr数据库中进行检索鉴定。最后用免疫组化和Western blotting方法验证差异蛋白LaminA/C在正常乳腺和乳腺癌组织的相对表达量。结果:经双向凝胶电泳和质谱鉴定,正常乳腺组含有（904 ± 58）个蛋白质点,乳腺癌组检测出含有（944 ± 70）个蛋白质点,共发现27个差异表达蛋白,成功鉴定出12种蛋白,其中包括LaminA/C。用免疫组化方法分析LaminA/C在正常乳腺和乳腺癌组织各20例中的表达,结果显示LaminA/C在正常乳腺和乳腺癌组织中的阳性率分别为15%（3/20）和40%（8/20）,P<0.05。用Western blot方法观察LaminA/C在正常乳腺和乳腺癌组织各10例中的表达,结果显示乳腺癌组中LaminA/C/β-actin 灰度值比值为0.40± 0.13,是正常乳腺组LaminA/C/β-actin 灰度值比值（0.21± 0.13）的1.91倍（P<0.05）。结论:LaminA/C在乳腺癌组织中表达明显增高,LaminA/C蛋白在乳腺癌的发生和发展中可能起一定的作用。      

Differential tissue-specific protein markers of vaginal carcinoma

The objective was to identify proteins differentially expressed in vaginal cancer to elucidate relevant cancer-related proteins. A total of 16 fresh-frozen tissue biopsies, consisting of 5 biopsies from normal vaginal epithelium, 6 from primary vaginal carcinomas and 5 from primary cervical carcinomas, were analysed using two-dimensional gel electrophoresis (2-DE) and MALDI-TOF mass spectrometry. Of the 43 proteins identified with significant alterations in protein expression between non-tumourous and tumourous tissue, 26 were upregulated and 17 were downregulated. Some were similarly altered in vaginal and cervical carcinoma, including cytoskeletal proteins, tumour suppressor proteins, oncoproteins implicated in apoptosis and proteins in the ubiquitin–proteasome pathway. Three proteins were uniquely altered in vaginal carcinoma (DDX48, erbB3-binding protein and biliverdin reductase) and five in cervical carcinoma (peroxiredoxin 2, annexin A2, sarcomeric tropomyosin kappa, human ribonuclease inhibitor and prolyl-4-hydrolase beta). The identified proteins imply involvement of multiple different cellular pathways in the carcinogenesis of vaginal carcinoma. Similar protein alterations were found between vaginal and cervical carcinoma suggesting common tumourigenesis. However, the expression level of some of these proteins markedly differs among the three tissue specimens indicating that they might be useful molecular markers.     

A reference map of human nasopharyngeal squamous carcinoma proteome

In order to conduct a comparative proteomics study of human nasopharyngeal carcinoma (NPC) to understand the molecular mechanisms that participate in the formation of NPC, the two-dimensional gel electrophoresis (2-DE) reference map of human NPC tissue proteome was described. To provide a high level of reproducibility between gels and accurately array each protein expressed in NPC tissue proteome, the two-dimensional polyacrylamide gel electrophoresis system, modified colloidal Coomassie Brilliant Blue staining method and ImageMaster 2D Platinum image analysis software were used. The NPC 2-DE maps show that high quality and good reproducibility of the 2-DE gel pattern was attained. An average total of 1,100 protein spots were separated by 2-DE, visualized by a modified colloidal Coomassie Brilliant Blue staining method. A synthesized 2-DE reference gel was acquired after detailed analysis of the NPC 2-DE gel maps, and 216 medium to high abundant spots were identified as landmark spots of NPC 2-DE gel, which expressed on >75% of gels. To provide an unambiguous identification of the landmark spots in gels, MALDI-TOF, ESI-Q-TOF mass spectrometry and database search were used to identify the proteins expressed in NPC tissue proteome. Between the 216 landmark spots, all proteins were identified with MALDI-TOF at first, 41 of which were identified with both MALDI-TOF and ESI-Q-TOF. All identified proteins were classified in terms of their subcellular localization and physiological function with information from SWISS-PROT and NCBI websites. According to our knowledge this is the first 2-DE reference map of human NPC. This reference map will serve as a basis for further studies of human NPC and the reference map data will be used to expand the proteome database of human NPC, which can be accessed in our website (http://www.xyproteomics.org/).     

高分化星形细胞瘤的蛋白质组学研究

目的 研究高分化星形细胞瘤差异蛋白质表达,为星形细胞瘤的治疗及预后的判断提供依据。方法 取经病理证实的29例正常脑组织及36例高分化的星形细胞瘤（Kernohan Ⅱ级）,经蛋白电泳、染色,采用PDQUEST和2-DE分析系统软件进行分析。以MALDI-TFO质谱或MALDI-TOF／TOF串联质谱技术结合数据库检索对蛋白质进行鉴定。结果 通过双向电泳得到正常脑组织和高分化星形细胞瘤标本的双向凝胶电泳图谱;生物质谱技术鉴定了24个差异蛋白质点,与正常脑组织相比,高分化星形细胞瘤有9个蛋白质下调,15个蛋白质上调。结论 以蛋白质组学技术鉴定了正常脑组织和高分化星形细胞瘤的差异蛋白质,其中部分蛋白质有助于深入研究星形细胞瘤的发生、发展机制并对进一步发现肿瘤标记物及治疗靶点有重要的参考价值。     

小细胞肺癌细胞系NCI-H446蛋白质表达谱的建立

背景与目的:小细胞肺癌(smallcelllungcancer,SCLC)是一种侵袭性极强的恶性肿瘤,具有增长迅速、早期转移等特点。目前公开的数据库中尚未见到小细胞肺癌的双向电泳参考图谱及其蛋白表达谱。本研究目的是建立高分辨率的小细胞肺癌细胞系NCI-H446细胞双向凝胶电泳图谱,并初步分析其蛋白质表达情况。方法:用固相pH梯度双向凝胶电泳技术(IPG-DALT)分离NCI-H446细胞总蛋白,凝胶银染显色,ImageMaster2D图像分析系统分析,从凝胶中选取分离较好的蛋白质点,应用基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)技术和数据库搜索鉴定蛋白质。结果:获得了背景清晰、分辨率和重复性较好的双向凝胶电泳图谱,三块胶平均蛋白质点数为1506±74,匹配点数为1412±56,匹配率达93.4%,三块胶蛋白质点在位置上有较好的重复性,不同胶间蛋白质点在IEF方向偏差是(0.96±0.27)mm,在SDS-PAGE方向偏差是(1.24±0.41)mm。胶内酶解-肽质指纹图分析鉴定了58个蛋白质,其中有原癌蛋白、细胞周期调控和信号转导相关蛋白质。结论:建立了小细胞肺癌细胞系NCI-H446双向电泳参考图谱,并应用质谱技术鉴定了部分蛋白质点,为进一步构建其蛋白质表达数据库提供了基础。     

Plasma proteomic analysis of patients with squamous cell carcinoma of the uterine cervix

Objective

To compare plasma protein expression between patients with squamous cell carcinoma (SCC) of the cervix and normal controls.

Methods

Plasma samples from patients with benign gynecological disease (normal cervix, n=6) and cervical cancer (SCC, n=6) were subjected to plasma proteomic analysis using two dimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization mass spectroscopy (MALDI-MS). Western blotting and immunoturbidimetric assay were performed to validate the results of 2-DE.

Results

Eight proteins showed differential expression between controls and SCC patients; six (ceruloplasmin, complement C3, afamin precursor, alpha-1-B-glycoprotein, transferrin, alpha-fibrinogen precursor) were up-regulated, while two (chain A, crystal structure of antithrombin and apolipoprotein A-IV precursor) were down-regulated in the plasma of SCC patients. Western blotting analysis revealed significant elevation of ceruloplasmin, complement C3, afamin, and alpha-1-B-glycoprotein in the plasma of SCC patients in comparison to controls. Immunoturbidimetric assay of a larger group confirmed the results of 2-DE and Western blotting, and showed that ceruloplasmin and complement C3 were significantly elevated in the plasma of SCC patients in comparison with controls and patients with carcinoma in situ (CIS) of the uterine cervix.

Conclusion

Plasma protein expression determined using 2-DE and MALDI-MS will give a chance to identify tumor-specific biomarkers for SCC of the cervix.     

维吾尔族妇女宫颈癌相关血浆候选肿瘤标志物的双向荧光差异电泳分析

目的:血清蛋白组学在肿瘤标志物的筛查研究中被广泛应用,本研究采用双向荧光差异电泳联合质谱技术,寻找与维吾尔族妇女宫颈鳞癌相关的血浆蛋白候选肿瘤标志物。方法:收集维吾尔族妇女宫颈病变血浆标本(慢性宫颈炎患者22例,宫颈癌早期患者26例),制备血浆低丰度蛋白质组样品,通过蛋白质双向荧光差异电泳技术分离与鉴别分析,筛选出慢性宫颈炎与宫颈癌早期患者两组间血浆差异表达的蛋白位点,运用质谱和生物信息学技术对其进行鉴定和功能识别分析。结果:确定了宫颈炎和早期宫颈鳞癌患者的血浆低丰度蛋白质组差异荧光电泳图谱,筛选出43个差异表达蛋白位点,通过质谱技术鉴定出16种蛋白质,在宫颈鳞癌早期患者血浆中有7个蛋白上调表达、9个蛋白下调表达。通过生物信息学分析,差异蛋白涉及的通路主要有补体和代谢相关蛋白和酶类。结论:本研究针对新疆维吾尔族妇女宫颈癌及官颈炎患者,展开血浆低丰度蛋白组表达水平的研究,鉴定出多种差异表达蛋白质,为宫颈癌的早期诊断及癌变机制研究提供了依据。     

结直肠癌化疗敏感性相关蛋白的蛋白质组学初步研究

目的:本研究应用蛋白质组学技术建立化疗敏感性不同的结直肠癌组织总蛋白双向凝胶电泳(two-dimensional gel electrophoresis,2-DE)图谱,并鉴定部分差异表达蛋白,以发现与结直肠癌化疗敏感性有关的蛋白.方法:收集临床诊断为晚期结直肠癌的病例,肠镜活检获取新鲜结直肠癌标本后液氮保存备用,根据肿瘤药物敏感实验分为化疗高敏感组和化疗低敏感组.提取组织总蛋白,采用双向凝胶电泳技术得到各组凝胶图谱;采用PD-quest 7.3软件进行图像的合成、对比和差异分析,识别化疗高敏感组和化疗低敏感组之间差异表达的蛋白斑点;选取差异蛋白质点,胶内酶解后行肽指纹图分析及网上数据库检索,鉴定差异蛋白质;应用Western印迹法检测部分差异蛋白的表达情况.结果: 建立了化疗高敏感组和化疗低敏感组的双向凝胶电泳图谱,多数蛋白质点集中于pH 4～8、相对分子质量为(20～100)×10~3.高敏感组和低敏感组的电泳图谱中平均蛋白质点数分别为(842±23)个和(793±19)个,2组平均匹配率为90.7%,2组间差异表达蛋白质点数为(79.00±13.56)个;选择30个差异蛋白质点进行质谱分析,经数据库查询鉴定出9个差异表达蛋白.结论:在化疗敏感性不同的结直肠癌中存在蛋白质表达的差异,这些差异表达蛋白可能与化疗敏感性有关,并可能用于化疗敏感性的预测.     

不同临床分期大肠癌组织的蛋白质组学比较研究

Objective: Colorectal carcinoma clinical stage associated proteins would be found by comparing differential expressed proteins from colorectal carcinoma tissues with different clinical stages. Methods: Total protein from colorectal carcinoma tissues were extracted; differential proteome profiles were established and analyzed by means of immobilized pH gradient-based two-dimensional polyacrylamide gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS). Results: Well-resolved, reproducible 2-DE profiles of human colorectal carcinoma tissues were obtained. Average protein spots were 970±41,980±32,1010±43,1240±34 in stage Ⅰ, stage Ⅱ,stage Ⅲ, stage Ⅳ respectively; Compared to stage Ⅰ, differential expressed protein spots was 52.00 ± 12 in stage Ⅱ, 42.00± 11 in stage Ⅲ, 72.00 ± 15 in stage Ⅳ; Part of differential expressing proteins were analyzed by mass spectrometry and bioinformation, 19 of them were well characterized. Three proteins were overexpressed in stage Ⅰ, stage Ⅲ, stage Ⅳ, and one protein were overexpressed in stage Ⅳ exclusively. Conclusion: Differential expressed proteins exist in clinical stage of colorectal carcinoma, which would be biomarkers for diagnosis and prediction of prognosis.     

双向凝胶电泳-飞行时间质谱法分析人肺鳞癌细胞NCI-H520蛋白质组成分

目的：建立和优化肿瘤蛋白质组研究的方法系统,并分析人肺鳞癌细胞蛋白质组。方法：用固相 pH梯度双向凝胶电泳分离人肺鳞癌细胞系 NCI H520总蛋白,银染显色, PDQuest 2DE软件分析,对部分蛋白质点用基质辅助激光解析电离飞行时间质谱 ( matrix assisted laser desorption/ionization time of flying mass spectrometry, MALDI TOF MS) 测定其胶内酶解后的肽质指纹图谱,用 PeptIdent软件查询 SWISS PROT数据库。结果：获得了分辨率和重复性均较好的双向电泳银染图谱,图像分析探测到 3块胶的平均蛋白质点数为 (1146± 116),平均匹配的点数为 (851± 95),匹配率达 73.7％, 3块胶在蛋白质点位置上具有较好的重复性, IEF方向的平均偏差为 (1.52± 0.22)mm, SDS PAGE方向的平均偏差为 (1.97± 0.13) mm。随机取 60个蛋白质点进行胶内原位酶解 - 质谱指纹图分析得到了 54个蛋白质点的肽质指纹图,查询数据库初步鉴定了 44个蛋白质,其中部分是与细胞周期有关的蛋白,部分是与信号传导有关的蛋白,部分是与癌基因相关的蛋白。结论：通过该研究建立了一套分辨率和重复性均较好的固向 pH梯度双向凝胶电泳分离细胞总蛋白和银染胶内原位酶解后 MALDI TOF MS肽质指纹图分析鉴定方法。通过该方法系统分析人肺鳞癌细胞 NCI H520蛋白质组成分而获得的资料将有益于人肺鳞癌细胞蛋白质组数据库的建立,从而为肺鳞癌的诊断、预防和治疗提供依据。     

人鼻咽癌细胞系HONE1的蛋白质双向凝胶电泳分析

目的：研究人鼻咽癌细胞系HONE1的蛋白质表达特征。方法：抽提鼻咽癌细胞系HONE1总蛋白,双向凝胶电泳（2－DE）分离、银杂显色,用凝胶成像仪和PDQUEST软件获取HONE1总蛋白的2－DE图像;根据获得的2－DE凝胶图像,从胶上切取分辨良好、染色较浓的蛋白质点,用胰蛋白酶原位水解,所获酶解肽段经基质辅助激光解吸飞行的时间质谱（MALDI－TOF－MS）测定Mr;以测得肽段Mr为肽质量指纹（PMF）,通过Pepldent软件搜索SWISS－PROT和TrEMBL数据库而识别该点对应的蛋白质。结果：建立了双向凝胶电泳分离总蛋白、质谱鉴定银染蛋白质点的方法,并鉴定出人鼻咽癌细胞系HONE1中表达较强的10个点相应的蛋白质。 结论：本研究为建立人鼻咽癌细胞HONE1的2－DE参考图和蛋白质数据库提供了有用的基础性研究资料,为进一步识别鼻咽癌特异性的蛋白质奠定了实验基础。     

Protein expression patterns in primary carcinoma of the vagina

Protein patterns in six samples from primary vaginal cancers, in five from normal vaginal tissue and in five primary cervical cancers, were analysed using two-dimensional polyacrylamide gel electrophoresis (2-DE). Protein expression profile was evaluated by computer-assisted image analysis (PDQUEST) and proteins were subsequently identified using matrix-assisted laser desorption/ionisation mass spectrometry. The aim was to analyse the protein expression profiles using the hierarchical clustering method in vaginal carcinoma and to compare them with the protein pattern in cervical carcinoma in order to find a helpful tool for correct classification and for increased biomedical knowledge. Protein expression data of a distinct set of 33 protein spots were differentially expressed. These differences were statistically significant (Mann-Whitney signed-Ranked Test, P<0.05) between normal tissue, vaginal and cervical cancer. Furthermore, protein profiles of pairs of primary vaginal and cervical cancers were found to be very similar. Some of the protein spots that have so far been identified include Tropomyosin 1, cytokeratin 5, 15 and 17, Apolipoprotein A1, Annexin V, Glutathione-S-transferase. Others are the stress-related proteins, calreticulin, HSP 27 and HSP 70. We conclude that cluster analysis of proteomics data allows accurate discrimination between normal vaginal mucosa, primary vaginal and primary cervical cancer. However, vaginal and cervical carcinomas also appear to be relatively homogeneous in their gene expression, indicating similar carcinogenic pathways. There might, further, be a possibility to identify tumour-specific markers among the proteins that are differentially expressed. The results from this study have to be confirmed by more comprehensive studies in the future.     

人Ⅰ期肺腺癌及癌旁组织蛋白质双向电泳图谱的差异分析

Objective: The aim of this study was to establish reproducible two-dimensional electrophoretic assay used for profiling and identification of differentially expressed proteins in human stage I lung adenocarcinoma and paired normal tu- mor-adjacent tissue. Methods: The proteins from 12 human stage I lung adenocarcinoma tissues and normal tumor-adjacent tissues were separated using isoelectric focusing electrophoresis (the first dimension) and the subsequent homogeneous SDS-polyacrylamide gel electrophoresis (SDS-PAGE) (the second dimension). The differentially expressed proteins were determined with PDQuest image analysis software, and identified using matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) and database searching. Results: The well-reproducible 2-DE gel patterns of human stage I lung adenocarcinoma and normal tumor-adjacent tissues were profiled and 26 differentially expressed proteins uncovered. Nine of these 26 protein spots were cut out from the preparation gels and determined with MALDI-TOF-MS. Searching against the protein database, four candidate proteins were identified. They were 60S acidic ribosomal protein P2, Cathepsin B1, Apolipoprotein A-I precursor, and La 4.1 protein. Conclusion: In this study, high reproducible 2-DE gel protein images of human stage I lung adenocarcinoma and paired normal tumor-adjacent tissues were achieved successfully, and 4 differentially expressed proteins were revealed. These data will be helpful for screen of early biomarker and study of molecular mechanisms of human lung adenocarcinoma.