【摘要】：目的通過(guò)差異蛋白質(zhì)組學(xué)研究,確定胰腺癌組織及相應(yīng)正常胰腺組織之間蛋白質(zhì)表達(dá)譜的差異,進(jìn)而闡明其表達(dá)水平的變化與胰腺癌發(fā)生發(fā)展之間的關(guān)系。方法選取胰腺癌組織及相應(yīng)正常胰腺組織6對(duì),提取總蛋白進(jìn)行雙向聚丙烯酰胺凝膠電泳,經(jīng)PDQuest軟件分析,獲得差異表達(dá)蛋白質(zhì)點(diǎn)。差異蛋白質(zhì)點(diǎn)經(jīng)過(guò)切取、酶解后進(jìn)行電噴霧四極桿飛行時(shí)間質(zhì)譜(ESI-Q-TOF MS/MS)鑒定。質(zhì)譜數(shù)據(jù)使用MASCOT搜索引擎對(duì)ExPASy蛋白質(zhì)序列數(shù)據(jù)庫(kù)進(jìn)行檢索,即獲得差異表達(dá)的蛋白質(zhì)。對(duì)鑒定出的部分蛋白質(zhì)通過(guò)免疫組化方法進(jìn)行驗(yàn)證。結(jié)果經(jīng)雙向凝膠電泳分析共有97個(gè)點(diǎn)被確定為差異表達(dá),其中31個(gè)點(diǎn)已被成功鑒定,23個(gè)蛋白上調(diào),8個(gè)蛋白下調(diào)。這些蛋白質(zhì)涉及體內(nèi)不同的功能過(guò)程和蛋白家族,主要包括:糖酵解相關(guān)酶,透明質(zhì)酸酶,Ⅰ、Ⅱ相代謝酶,金屬蛋白酶,細(xì)胞骨架,Ca2+代謝相關(guān)酶等。免疫組化方法進(jìn)一步證實(shí)了HYAL1表達(dá)上調(diào),CYP2C8和GSTM2表達(dá)下調(diào)。結(jié)論應(yīng)用比較蛋白質(zhì)組學(xué)方法可篩選出胰腺癌患者癌組織和正常胰腺組織間存在的差異表達(dá)蛋白,篩選出的蛋白將為進(jìn)一步闡述胰腺癌的發(fā)生發(fā)展機(jī)制提供很好的線索和幫助。
[Abstract]:Objective to determine the difference of protein expression profiles between pancreatic cancer tissues and normal pancreatic tissues by differential proteomics, and to elucidate the relationship between the changes of protein expression levels and the occurrence and development of pancreatic cancer. Methods six pairs of pancreatic cancer tissues and corresponding normal pancreatic tissues were selected and total proteins were extracted by two-dimensional polyacrylamide gel electrophoresis. The differentially expressed protein spots were obtained by PDQuest software analysis. The differential protein spots were digested and hydrolyzed by electrospray quadrupole mass spectrometry (ESI-Q-TOF MS/MS). The MASCOT search engine was used to search the ExPASy protein sequence database and the differentially expressed proteins were obtained. Some of the identified proteins were verified by immunohistochemical method. Results A total of 97 sites were identified as differentially expressed by two-dimensional gel electrophoresis, 31 of which were successfully identified, 23 proteins were up-regulated and 8 proteins were down-regulated. These proteins are involved in different functional processes and protein families in vivo, including glycolytic enzymes, hyaluronidase, phase 鈪，

本文編號(hào)：2312383