CAJ | 학술논문

目的了解负载不同转移潜能肝癌细胞裂解物和正常肝细胞裂解物后树突状细胞蛋白质表达谱的差异.探索肝癌转移机制.方法以不同转移潜能人肝癌细胞株(HCCLM6-高转移、MHCC97L-低转移、Hep3B-不转移)制备裂解物与正常肝细胞株(ChangLiver)制备裂解物分别负载DC,并以无裂解物负载DC为对照,对DC全蛋白进行双向电泳、银染、电喷雾质谱分析,Pdquest软件查询IPI上人的非冗余蛋白数据库鉴定差异蛋白.结果双向电泳图谱匹配率>80%,相关因子平均>0.8;发现最大和最小差异大于3倍的蛋白质点21个,初步鉴定其中的12个.发现这些蛋白涉及细胞骨架(3号点:Ⅰ型角蛋白CK10;16号点:TPMsk3家族蛋白;21号点:β-中心体肌动蛋白)、信号传导(8号点:钙黏蛋白家族成员;11号点:膜联蛋白A2异构体;17号点:内涵体/溶酶体MP1作用蛋白前体)、细胞动力和能量代谢(6号点:细胞色素辅酶Q还原酶复合体核心蛋白;9号点:胆绿素还原酶A前体;13号点:苹果酸脱氢酶;20号点:线粒体前体)等方面及一些未归类蛋白(10号点:抗结直肠癌重链;18号点:Transgelin2).结论负载不同转移潜能肝癌裂解物后DC蛋白质谱的变化涉及细胞骨架、信号传导、细胞质内物质运输和能量代谢等方面.其中部分蛋白可能与DC2功能和肝癌转移潜能相关.
목적 료해부재불동전이잠능간암세포렬해물화정상간세포렬해물후수돌상세포단백질표체보적차이.탐색간암전이궤제.방법 이불동전이잠능인간암세포주(HCCLM6-고전이、MHCC97L-저전이、Hep3B-불전이)제비렬해물여정상간세포주(ChangLiver)제비렬해물분별부재DC,병이무렬해물부재DC위대조,대DC전단백진행쌍향전영、은염、전분무질보분석,Pdquest연건사순IPI상인적비용여단백수거고감정차이단백.결과 쌍향전영도보필배솔>80%,상관인자평균>0.8;발현최대화최소차이대우3배적단백질점21개,초보감정기중적12개.발현저사단백섭급세포골가(3호점:Ⅰ형각단백CK10;16호점:TPMsk3가족단백;21호점:β-중심체기동단백)、신호전도(8호점:개점단백가족성원;11호점:막련단백A2이구체;17호점:내함체/용매체MP1작용단백전체)、세포동력화능량대사(6호점:세포색소보매Q환원매복합체핵심단백;9호점:담록소환원매A전체;13호점:평과산탈경매;20호점:선립체전체)등방면급일사미귀류단백(10호점:항결직장암중련;18호점:Transgelin2).결론 부재불동전이잠능간암렬해물후DC단백질보적변화섭급세포골가、신호전도、세포질내물질운수화능량대사등방면.기중부분단백가능여DC2공능화간암전이잠능상관.
Objective To explore the mechanism of HCC immune escape, we utilized proteomic approach to analyze the relations between expression profiles changes of dendritic cells (DCs) pulsed with hepatocellular carcinoma (HCC) lysates of different metastatic potentials and the changes of DC's function. Methods DCs were derived from human peripheral blood monocytes (PBMC) and pulsed with HCC lysates of different metastatic potentials. The normal liver cell line lysate-pulsed DCs and norrlysate pulsed DCs were employed to serve as the control. The total protein of these DCs was identified by immobilized pH gradient two-dimensional poly-acrylamide gel electrophoresis (2DE), silver stain, Pdquest 2DE analysis software, ESI, and IPI database search to yield the different profiles among them. Results The match ratio of all samples' 2DE was higher than 80% while the correlation coefficient over 0. 8. There were 21 spots exhibiting quantitative changes that more than 3 folds among DCs pulsed with different lysates, 12 of them were identified by ESI. These proteins were related to cytoskeleton ( spot 3: Keratin type 1 cytoskeletal 10 ; spot 16: TPMsK3; and spot 21: p-centractin), signal transduction ( spot 8: Cadherin-related tumor suppressor homolog precersor; spot 11 Annexin A2 Iso-form 2; spot 17: Late endosomal/Lysosomal Mp 1 interacting precursor), cytoplasmic movement and energic metabolism ( spot 6:Ubiquinol-cytochrome C reductase complex core protein I, mitochondrial precursor;spot 9: Biliverdin reductase A precersor ; spot 13: Cytosolic malate dehydrogenase ; and spot 20: Methylmalonate-semialdehyde dehydrogenase [cylating], mitochondrial precursor) and others ( spot 10: Anti-colorectal carcinoma heavy chain ;spot 18: Transgelin-2 ). Conclusion These 12 spots represent proteins of cytoskeleton, signal transduction as well as proteins involved in cytoplasmic movement and energic metabolism. Some of them might be related to the function of DCs and metastatic potential of HCC.