肿瘤研究与临床
腫瘤研究與臨床
종류연구여림상
Cancer Research and Clinic
2015年
9期
586-588,592
,共4页
柴丽丽%陈燕%王娟红%郑长黎
柴麗麗%陳燕%王娟紅%鄭長黎
시려려%진연%왕연홍%정장려
胃肿瘤%腺癌%淋巴转移%显微切割%光谱法,质量,基质辅助激光解吸电离%蛋白质组学
胃腫瘤%腺癌%淋巴轉移%顯微切割%光譜法,質量,基質輔助激光解吸電離%蛋白質組學
위종류%선암%림파전이%현미절할%광보법,질량,기질보조격광해흡전리%단백질조학
Stomach neoplasms%Adenocarcinoma%Lymphatic metastasis%Microdissection%Spectrometry,mass,matrix-assisted laser desorption-ionization%Proteomics
目的：利用比较蛋白质组学技术识别并鉴定胃癌原发灶与淋巴结转移灶中的差异表达蛋白质，筛选特异性的转移相关蛋白，探讨胃癌淋巴结转移的分子机制。方法将前期应用二维差异凝胶电泳技术（DIGE）获取人胃癌原发灶及转移灶差异蛋白表达图谱进行分析获得的11个差异表达蛋白质，运用基质辅助激光解吸电离飞行时间质谱（MALDI-TOF-MS）进行鉴定，得到相应的肽质量指纹图（PMF），然后搜索数据库鉴定出部分蛋白质。利用免疫组织化学技术检测HSP 70在胃癌中的表达水平。结果对11个差异蛋白质点进行PMF分析后，通过Mascot软件查询鉴定出其中5个蛋白质点（HSP70的8同种型2变异体、2个伴侣蛋白、亮氨酸氨肽酶、预测假想蛋白XP_515584）。 HSP70、伴侣蛋白、亮氨酸氨肽酶的表达在原发癌灶中明显升高。免疫组织化学检测示HSP 70的表达水平随着胃癌恶性程度的增加及淋巴结的转移逐渐增加，与蛋白质组学的分析结果一致。结论人胃癌原发灶与淋巴结转移灶中腺癌细胞的差异蛋白表达图谱相似，说明两者癌细胞的来源、分化程度相对一致，仅有少数蛋白质的质和量发生了明显变化，参与了淋巴结转移。其涉及细胞生理活动的各方面，与肿瘤的发生、发展等密切相关。 HSP70参与了胃癌的发生发展，其表达水平与胃癌有无淋巴结转移及恶性程度有密切关系。
目的：利用比較蛋白質組學技術識彆併鑒定胃癌原髮竈與淋巴結轉移竈中的差異錶達蛋白質，篩選特異性的轉移相關蛋白，探討胃癌淋巴結轉移的分子機製。方法將前期應用二維差異凝膠電泳技術（DIGE）穫取人胃癌原髮竈及轉移竈差異蛋白錶達圖譜進行分析穫得的11箇差異錶達蛋白質，運用基質輔助激光解吸電離飛行時間質譜（MALDI-TOF-MS）進行鑒定，得到相應的肽質量指紋圖（PMF），然後搜索數據庫鑒定齣部分蛋白質。利用免疫組織化學技術檢測HSP 70在胃癌中的錶達水平。結果對11箇差異蛋白質點進行PMF分析後，通過Mascot軟件查詢鑒定齣其中5箇蛋白質點（HSP70的8同種型2變異體、2箇伴侶蛋白、亮氨痠氨肽酶、預測假想蛋白XP_515584）。 HSP70、伴侶蛋白、亮氨痠氨肽酶的錶達在原髮癌竈中明顯升高。免疫組織化學檢測示HSP 70的錶達水平隨著胃癌噁性程度的增加及淋巴結的轉移逐漸增加，與蛋白質組學的分析結果一緻。結論人胃癌原髮竈與淋巴結轉移竈中腺癌細胞的差異蛋白錶達圖譜相似，說明兩者癌細胞的來源、分化程度相對一緻，僅有少數蛋白質的質和量髮生瞭明顯變化，參與瞭淋巴結轉移。其涉及細胞生理活動的各方麵，與腫瘤的髮生、髮展等密切相關。 HSP70參與瞭胃癌的髮生髮展，其錶達水平與胃癌有無淋巴結轉移及噁性程度有密切關繫。
목적：이용비교단백질조학기술식별병감정위암원발조여림파결전이조중적차이표체단백질，사선특이성적전이상관단백，탐토위암림파결전이적분자궤제。방법장전기응용이유차이응효전영기술（DIGE）획취인위암원발조급전이조차이단백표체도보진행분석획득적11개차이표체단백질，운용기질보조격광해흡전리비행시간질보（MALDI-TOF-MS）진행감정，득도상응적태질량지문도（PMF），연후수색수거고감정출부분단백질。이용면역조직화학기술검측HSP 70재위암중적표체수평。결과대11개차이단백질점진행PMF분석후，통과Mascot연건사순감정출기중5개단백질점（HSP70적8동충형2변이체、2개반려단백、량안산안태매、예측가상단백XP_515584）。 HSP70、반려단백、량안산안태매적표체재원발암조중명현승고。면역조직화학검측시HSP 70적표체수평수착위암악성정도적증가급림파결적전이축점증가，여단백질조학적분석결과일치。결론인위암원발조여림파결전이조중선암세포적차이단백표체도보상사，설명량자암세포적래원、분화정도상대일치，부유소수단백질적질화량발생료명현변화，삼여료림파결전이。기섭급세포생리활동적각방면，여종류적발생、발전등밀절상관。 HSP70삼여료위암적발생발전，기표체수평여위암유무림파결전이급악성정도유밀절관계。
Objective To identify the differentially-expressed proteins of human gastric adenocarcinoma cell in primary or metastatic lymph node tissues by comparative proteomics technology, and to screen the specific metastatic-associated proteins so as to investigate the metastatic molecular mechanism of lymph node metastasis in gastric cancer. Methods 11differential proteins were acquired previously from primary and metastatic lymphnode tissues in gastric adenocarcinoma patients by 2D-DIGE. Some selected differential protein spots were identified by PMF based on MALDI-TOF-MS and database search. Immunohistochemical staining of HSP70 was used to evaluate the reliability of the proteomic analysis results. Results After analyzed on 11 differential proteins by in-gel trypsin digestion and MALDI-TOF-MS-based PMF analysis, a total of 5 differential proteins were identified by searching Mascot-database, HSP70ˊs 8 isoform 2 variant, chaperonin, chaperonin, leucine aminopeptidase, predicted: hypothetical protein XP_515584. Among the differential proteins identified, the levels of HSP70, chaperonin, leucine aminopeptidase expression had a significant up-regulation in gastric primary cancer compared with metastatic lymph node. HSP70 expression rate increased with the metastasis of lymph node and the progress of gastric cancer, agreed with the proteomics results. Conclusions They are similar in differentially-expressed proteins in primary or metastatic lymph node tissues because of the uniformity in source and differention. There are few protein changes in cancer cells between them, taking part in the metastatic of gastric cancer. HSP70 takes part in the progress of gastric cancer and relates to the metastasis of lymph node and malignant degree.
