CAJ | 학술논문

目的应用比较蛋白质组学技术对不同分化程度胃癌组织的差异蛋白质进行筛选鉴定.方法十二烷基硫酸钠聚丙烯酰氨凝胶电泳(SDS-PAGE)结合液质联用(LC-MS)技术对8例不同分化胃腺癌组织的差异蛋白质进行鉴定,并应用实时荧光定量核酸扩增检测系统(QPCR)和Western blot法对鉴定出的部分因子进行验证.结果差异表达明显的蛋白质电泳条带15条,共鉴定出355种蛋白质,筛选后经功能分析确定8种可能与肿瘤分化有关的蛋白质,为人肌动蛋白素(Cofilin)、蛋白酶体激活亚单位1(PSME1)、Ras相关蛋白7a(RAB7a)、细丝蛋白A(FLNa)、丙酮酸激酶(PK)、谷胱甘肽巯基转移酶-π-1( GST-π-1)、过氧化物酶5(PRDX5)、actin相关蛋白.对Cofliin、Ras相关蛋白7a、PSME1进行验证,证实这些因子的表达与蛋白质组学鉴定的结果相符.结论分化程度不同的胃腺癌组织蛋白质表达存在差异.
목적 응용비교단백질조학기술대불동분화정도위암조직적차이단백질진행사선감정.방법 십이완기류산납취병희선안응효전영(SDS-PAGE)결합액질련용(LC-MS)기술대8례불동분화위선암조직적차이단백질진행감정,병응용실시형광정량핵산확증검측계통(QPCR)화Western blot법대감정출적부분인자진행험증.결과 차이표체명현적단백질전영조대15조,공감정출355충단백질,사선후경공능분석학정8충가능여종류분화유관적단백질,위인기동단백소(Cofilin)、단백매체격활아단위1(PSME1)、Ras상관단백7a(RAB7a)、세사단백A(FLNa)、병동산격매(PK)、곡광감태구기전이매-π-1( GST-π-1)、과양화물매5(PRDX5)、actin상관단백.대Cofliin、Ras상관단백7a、PSME1진행험증,증실저사인자적표체여단백질조학감정적결과상부.결론 분화정도불동적위선암조직단백질표체존재차이.
Objective To identify differentiation-related proteins in gastric adenocarcinoma by comparative proteomics.Methods The holoproteins of human gastric adenocarcinoma tissues in 4 well differentiated and 4 poorly differentiated were measured by sodium dodecyl-sulphate (SDS) electrophoresis and liquid chromatography-mass spectrometry (LC-MS),and differentiation-related proteins in gastric adenocarcinoma were analyzed.Some proteins obtained through proteomics were tested by Real-time quantitative polymerase chain reaction detecting system (QPCR) and Western blotting in the tissues of gastric carcinoma.Results Fifteen differential protein bands were found in gastric adenocarcinoma,among them 355 proteins were identified by LC-MS.The proteins such as high abundant proteins and structural proteins were ruled out,then 8 proteins were selected and these may be meaningful,which included cofilin,proteasome activator subunitl,ras-related protein rab-7a,filamin a,pyruvate kinase,glutathione s-transferasesπ-1,peroxiredoxin 5,actin-related protein.These proteins are involved in the signal transduction,maintenance of cell structure,and regulate cell proliferation and apoptosis.The expression of 3 proteins identified by Western blotting and QPCR was consistent with the results obtained by proteomics.Conclusion Differential proteins are found in gastric adenocarcinoma tissues with different differentiation degrees.