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1.
目的:建立肝癌血清学诊断模型,探讨评估SELDI-TOF-MS技术在肝癌诊断和介入治疗评价中的价值.方法:用弱阳离子交换芯片(CM10芯片)和表面增强激光解吸电离飞行时间质谱仪(surface-enhanced laser desorption ionization time-of-flight mass spectrometry,SELDI-TOF-MS)技术,测定60例肝癌患者和60例正常对照者的血清蛋白质指纹图谱,应用BiomarkerWizard统计软件比较肝癌组和正常对照组血清蛋白质表达的差异性,采用Biomarker Pattern软件分析数据建立肝癌诊断模型,比较介入治疗前后血清蛋白质指纹图谱的差异性.结果:在质荷比(M/Z)为2000-10000范围内,和正常血清比较,肝癌的差异峰有3个(M/Z为4182Da、5710Da、6992Da;P<0.01),4182Da和5710Da下调,6992Da上调.用这3个差异蛋白峰建立肝癌诊断模型,诊断肝癌的灵敏度为93.3%(28/30),特异度为90.0%(27/30),正确率为91.7%(55/60),约登指数为0.833.差异蛋白峰(M/Z4182Da)在介入术后1mo明显上调(P<0.05).结论:应用SELDI-TOF-MS技术进行肝癌血清蛋白质指纹图谱分析,建立肝癌诊断树模型,对肝癌的诊断有一定的价值;筛选出的差异蛋白峰对肝癌的介入治疗评估有一定的应用价值.  相似文献   

2.
应用SELDI-TOF-MS技术建立肝癌筛选血清蛋白质指纹图谱模型   总被引:8,自引:0,他引:8  
目的:建立肝癌筛选血清蛋白质指纹图谱模型.方法:用表面加强激光解析电离飞行时间质谱技术(SELDI-TOF-MS)及WCX2蛋白芯片获得新发肝癌、肝硬化患者和正常人血清的蛋白质指纹图谱,用计算机软件进行比较分析,建立肝癌的筛选模型.结果:肝癌患者与健康对照组血清蛋白质指纹图谱之间有5个标志蛋白(4477,8943,5181, 8617,13 761 Da)在肝癌患者血清中高表达,肝癌患者与肝硬化患者血清蛋白质指纹图谱之间2个标志蛋白(4477,13 761 Da)在肝癌患者血清中高表达,1个标志蛋白(4097 Da)在肝癌患者血清中低表达.SELDI-TOF-MS技术的特异性(60/60,100%);敏感度(18/20,90%).分析系统筛选出4477,8943,13 761,4097 Da标志蛋白建立的肝癌诊断模型.结论:建立的血清蛋白质指纹图谱模型能够区分肝癌与非肝癌患者,SELDI-TOF-MS在肝癌的诊断及肿瘤特异性蛋白质生物标志分子的筛选等方面具有一定价值.  相似文献   

3.
王瑜  余力  丁彦青  刘莉  肖莎 《山东医药》2011,51(32):10-12,118
目的观察CD24、CD73、Hsp27和Fascin蛋白在大肠癌组织中的表达,探讨其与大肠癌亲器官转移的相关性及临床病理意义。方法应用免疫组织化学技术检测Fascin、CD24、CD73和Hsp27蛋白在20例无转移大肠癌和19例同时伴有淋巴结和肝脏转移的大肠癌的不同部位癌组织中的表达。结果转移性大肠癌原发灶与未转移大肠癌比较,CD73、Fascin的表达差异显著(P=0.009,0.009),Hsp27、CD24的表达无明显差异。CD24在淋巴结转移灶的表达与肝转移灶和原发灶比较,差异显著(P=0.000,0.001);原发灶和肝转移灶的表达比较,差异不显著。CD73肝转移灶的表达与淋巴结转移灶和原发灶比较差异显著(P=0.003,0.000);而淋巴结转移灶和原发灶的表达比较无明显差异。Hsp27在肝转移灶的表达与原发灶比较,无明显差异;与淋巴结转移灶比较,差异显著(P=0.001);在原发灶与淋巴结转移灶的表达比较差异显著(P=0.021)。Fascin在原发灶的表达与肝转移灶、淋巴结转移灶比较无明显差异。结论 CD24与大肠癌淋巴结转移相关,可能是大肠癌淋巴结转移的预测因子之一;CD73、Hsp27的高表达与大肠癌肝转移有关,Fascin的表达与大肠癌转移有关,但与大肠癌转移至淋巴结或肝脏无相关性。  相似文献   

4.
目的:应用表面增强激光解吸附电离飞行时间质谱技术(SELDI-TOF-MS)对肝细胞癌组织的蛋白表达谱进行分析,从中筛选出标记蛋白.方法:采用CM10芯片及SELDI-TOF-MS技术对肝细胞癌组织26例和肝硬化组织18例进行蛋白指纹图谱检测分析;比较高、中、低分化肝细胞癌组织,不同TNM分期肿瘤以及AFP阴、阳性肝细胞癌组织的蛋白表达差异,所得结果均采用Biomarker Wizard软件进行分析;通过查询蛋白库,对特定分子质量所对应的标记蛋白进行初步确定.结果:肝细胞癌和肝硬化组织蛋白表达图谱之间存在16个稳定的标志蛋白,7个蛋白在肝细胞癌组织中表达上调.9个蛋白在肝细胞癌组织中表达下调,其中4.7 kDa,7.2 kDa和9.8 kDa蛋白峰差异性最明显;中分化和高分化肝细胞癌之间存在11个标志蛋白;通过查询蛋白库ExPasy并进行筛选,初步确定特定分子质量所对应的蛋白.结论:采用SELDI-TOF-MS技术,证实在肝细胞癌组织中存在多种高度特异性低分子蛋白.  相似文献   

5.
SELDI技术筛选肺癌患者血清标志蛋白的临床价值   总被引:2,自引:0,他引:2  
目的探讨表面增强激光解析电离飞行时间质谱(SELDI-TOF-MS)技术筛选肺癌患者血清标志蛋白的临床价值。方法用SELDI-TOF-MS技术、弱阳离子交换蛋白芯片,检测肺癌和肺良性病变患者的血清蛋白质质谱图;用Biomarker Pattern软件分析肺癌差异蛋白并初建其诊断模型,通过盲筛验证诊断模型。结果发现有统计学差异的蛋白峰20个,其中肺癌患者血清高表达蛋白质波峰14个,低表达蛋白质波峰6个;用质荷比2 090.77、2 503.31 Da的差异蛋白峰建立分类树模型,其诊断肺癌的灵敏度88%,特异度95%;盲筛验证灵敏度90%,特异度100%,粗符合率93.33%,Youden指数0.9。结论SELDI-TOF-MS技术筛选的肺癌血清差异性蛋白及分类树模型,诊断肺癌的灵敏度高、特异性好。  相似文献   

6.
目的:检测HBV感染携带者与肝细胞癌(HCC)患者血清蛋白质的差异性表达,以发现HCC的肿瘤诊断标志物.方法:用表面增强的激光解析电离飞行时间质谱(SELDI-TOF-MS)技术检测27例HCC患者,27例HBV感染携带者,25例健康对照血清中的蛋白质谱,并用Biomarker Patterns System 5.0软件分析,建立诊断模型.结果:检测HCC患者与HBV感染携带者。正常对照与HCC患者,正常对照与HBV感染携带者的差异性蛋白分子,据此构建分类模型,得到的灵敏度和特异度分别为93%,85%; 96%,96%;84%,89%.其中相对分子质量为8141 Da的蛋白峰在HCC组明显高于HBV感染组(p<10-5);相对分子质量为3448 Da的蛋白峰在HCC及HBV感染携带组表达均较正常组显著增高(P<10-5),而在HCC-HBV组无明显差异(P>0.05),提示其可能为HBV感染的一种标志蛋白;相对分子质量为777) Da的蛋白峰在3组中均有差异性表达.结论:SELDI蛋白芯片技术检测血清蛋白质谱法诊断HCC具有较高的灵敏度和特异度,操作简单快捷,临床应用前景广阔,为HCC诊断提供了新的血清学方法.  相似文献   

7.
目的:分析肝内胆管癌细胞系ICC-9810与正常肝细胞系L02的蛋白质表达差异,筛选肝内胆管癌的潜在分子标志物.方法:应用表面增强激光解吸离子化(surface enhanced laser desorption/ionization,SELDI)蛋白质芯片技术检测肝内胆管癌及正常肝细胞系的蛋白质谱.用PBSII-C型蛋白质芯片阅读机读取数据,采用Proteinchip Software 3.0.2软件分析数据.结果:IMAC3、WCX2两种蛋白芯片共捕获376个蛋白峰,发现27个差异蛋白.与正常肝细胞系L02蛋白谱相比,9个蛋白在肝内胆管癌细胞系ICC-981O中高表达,18个蛋白在肝内胆管癌细胞系ICC-9810中低表达.其中3767、7999、10555、12163、22066和26794 Da6个差异蛋白可被WCX2和IMAC3芯片共同捕获.结论:肝内胆管癌与正常肝细胞存在差异蛋白表达,这些差异蛋白为寻找肝肿瘤标志物,了解肝内胆管癌的发病机制提供了重要线索.  相似文献   

8.
目的采用表面增强激光解析电离飞行时间质谱(SELDI-TOF-MS)技术筛选双侧肾母细胞瘤(BWT)的差异性表达蛋白。方法选取BWT患儿血清标本10例(BWT组),另选单侧肾母细胞瘤(UWT)患儿血清标本(UWT组)及健康儿童血清标本(对照组)各20例,采用SELDI-TOF-MS技术检测并收集相关数据。结果应用生物信息学方法对所得数据进行分析,得到差异性峰10个(P<0.01)。对比各组差异性表达蛋白峰值数据,筛选出质子数/电荷数(m/z)为5 648 Da的差异性表达蛋白1个,其在BWT组、UWT组及对照组中表达强度分别为3 889.36±1 796.83、2 886.81±1 404.65、432.21±730.42,三组相比,P均<0.01。结论采用SELDI-TOF-MS技术成功筛选出一种BWT差异性表达蛋白,其有望成为BWT早期诊断及预后判断的新的标志物。  相似文献   

9.
大肠癌发展与肝转移的差异蛋白质组研究进展   总被引:2,自引:2,他引:0  
近年来大量统计资料表明,大肠癌作为临床最常见的恶性肿瘤之一,其发病率和死亡率逐年增高.肝脏是大肠癌最常见、最易发生的转移器官.肝转移是大肠癌根治性切除后死亡的主要原因.所以,早期预测和诊断肝转移对于提高大肠癌患者生存率,改善预后有重要意义.差异蛋白质组研究(又称功能蛋白质组研究) 利用蛋白质组研究技术,对正常大肠黏膜组织、大肠癌原发灶、大肠癌邻近组织、大肠癌肝转移灶、血浆或血液、组织液和尿液进行蛋白分子代谢产物的定性和定量分析并与正常人蛋白代谢谱进行对照分析,可以发现大肠癌中特有的小分子代谢物,进一步探讨大肠癌肝转移的发生机制,观察由多基因事件引起的多蛋白质组分整体变化,从整体上寻找潜在的药物靶点并且通过肿瘤标志物进行大肠癌的早期诊断和治疗,防止其发展与转移.我们主要就大肠癌发展与肝转移的差异蛋白质组研究的理论、方法以及成果进行了初步的回顾和总结.  相似文献   

10.
目的研究半胱氨酸天冬氨酸蛋白酶3(caspase-3)表达在老年贲门癌发生发展中的作用。方法采用免疫组化S-P法观察113例老年贲门癌旁上皮细胞、原发癌细胞和浸润淋巴细胞以及转移灶癌细胞中caspase-3表达,并分析原发灶癌细胞caspase-3表达与贲门癌临床病理特征的关系。结果caspase-3在贲门癌旁上皮细胞中的表达高于原发灶癌细胞(P<0·05);caspase-3在贲门癌原发灶浸润淋巴细胞中的阳性率高于贲门癌旁上皮细胞和原发灶癌细胞(P<0·05);转移灶癌细胞中caspase-3阳性率高于原发灶癌细胞(P<0·05);原发灶癌细胞中caspase-3表达与贲门癌患者的年龄、性别以及贲门癌肿瘤大小、浸润深度、转移、TNM分期、生长方式和分化程度无相关性(P<0·05)。结论贲门癌原发灶癌细胞caspase-3表达下调和浸润淋巴细胞caspase-3表达上调与贲门癌发生有关,来自原发灶和转移微环境中的化学物质可能通过提高转移灶癌细胞中caspase-3表达,进而起到抑制转移灶的作用。  相似文献   

11.
AIM: To identify the differential proteins associated with colorectal cancer genesis and hepatic metastasis.METHODS: Hydrophobic protein samples were extracted from normal colorectal mucosa, primary cancer lesion and hepatic metastatic foci of colorectal cancer. With twodimensional electrophoresis and image analysis,differentially expressed protein spots were detected, and the proteins were identified by matrix assisted laser desorption/ionization-time of flight-mass spectrometry and peptide mass fingerprint analysis.RESULTS: Significant alterations of the proteins in number and expression levels were discovered in primary cancer and hepatic metastatic foci, the expression of a number of proteins was lost in 25-40 ku, but protein spots was increased in 14-21ku, compared with normal mucosa. Nine differentially expressed protein spots were identified. Three proteins expressed in normal mucosa, but lost in primary cancer and hepatic metastasis, were recognized as calmodulin, ribonuclease 6 precursor and mannosidase-α.Proapolipoprotein was expressed progressively from normal mucosa to primary cancer and hepatic metastasis. The differentially expressed protein of beta-globin was found in normal mucosa and hepatic metastatic tumor, but lost in primary cancer lesion. Cdc 42, a GTP-binding protein, was identified in hepatic metastasis. The protein spots of C4 from primary cancer, M7 and M9 from hepatic metastasis had less homology with the proteins in database.CONCLUSION: Variations of hydrophobic protein expression in colorectal cancer initiation and hepatic metastasis are significant and can be observed with two-dimensional electrophoresis. The expression of calmodulin, ribonuclease 6 precursor and mannosidase-α is lost but the expression of proapolipoprotein is enhanced which is associated with colorectal cancer genesis and hepatic metastasis. Cdc 42 and beta-globin are expressed abnormally in hepatic metastasis. Protein C4, M7 and M9 may be associated with colorectal cancer genesis and hepatic metastasis.  相似文献   

12.
目的:研究遗传性非息肉病性结直肠癌(HNPCC)与散发性结直肠癌患者术前血清蛋白质的表达差异,以期发现可用于HNPCC诊断的生物学指标.方法:应用表面增强激光解吸电离飞行时间质谱(SELDI-TOF-MS)技术结合蛋白质芯片分别检测20例HNPCC和25例散发性结直肠癌患者术前血清蛋白质组分.将获得的蛋白质谱采用美国C...  相似文献   

13.
To clarify the relationship between multidrug resistance protein (MRP) and clinicopathologic features, the influence of adjuvant chemotherapy, and prognosis of patients who underwent resection of metastatic liver carcinomas originating from colorectal carcinomas, we examined the expression of MRP in tumor tissues by immunostaining. Specimens of 38 primary colorectal tumors and 44 metastatic liver tumors of colorectal origin were examined (metastatic group). We also examined 28 nonmetastatic colorectal carcinomas. The percentages of nonmetastatic tumors and of primary and metastatic tumors of the metastasis group that expressed MRP were similar. MRP expression in primary and metastatic tumors did not correlate with any clinicopathologic features. The use of adjuvant chemotherapy after operation for primary colorectal carcinomas was associated with increased MRP expression among metastatic liver tumors. Expression of MRP in the tumor did not influence the prognosis or survival rate after resection of primary or metastatic tumors. Our data suggest that MRP expression in metachronous liver metastases from colorectal carcinomas may be induced by administration of anticancer drugs but is not associated with clinicopathologic features of the tumor, liver metastasis, or prognosis. Received: November 19, 1998 / Accepted: May 28, 1999  相似文献   

14.
目的:分析结直肠癌(colorectal cancer,CRC)与正常黏膜间的蛋白质表达差异,筛选新的肿瘤标志物;并对兴趣蛋白进行验证,分析其与CRC的发生、发展及淋巴结转移的关系.方法:对6对新鲜的CRC与正常黏膜组织进行以二维差异凝胶电泳(2D differential gel electrophoresis,2D DIGE)及基质辅助激光解吸飞行时间质谱(matrix-assisted laserde sorption/ionization-time of flight masss pectrometry,MALDI-TOF-MS)分析.以免疫组织化学法验证兴趣蛋白泛醌细胞色素c还原酶核心蛋白1(ubiquinol cytochrome-creductase core protein 1,UQCRC1)在78例CRC与正常黏膜组织,和24个转移淋巴结中的表达.对染色的强弱评分为阴性:0,弱阳性:1,中阳性:2,强阳性:3.结果:2DDIGE分析显示在CRC中一个蛋白点丰度平均显著增高2.14倍(P<0.001).MALDI-TOF-MS分析证实该蛋白为UQCRC1.免疫组织化学法分析显示UQCRC1在CRC与正常黏膜组织中表达分别为2.28±0.95和1.81±0.88,有显著差异(P<0.001).UQCRC1表达的强弱与分化、分期及部位均无关(P>0.05).UQCRC1在转移淋巴结与配对的原发灶中表达分别为2.79±0.51和2.33±0.96,有显著差异(P<0.05).结论:UQCRC1在结直肠癌变和淋巴结转移过程中发挥一定的作用.  相似文献   

15.
李杰  陈茂伟  周怡  邹俊 《内科》2011,6(1):9-11,99
目的探讨早期病毒性乙型肝炎(乙肝)肝纤维化的潜在血清标志物并建立相应的诊断模型。方法采用表面增强激光解吸/离子化时间飞行质谱(SELDI-TOF-MS)技术检测20个健康人和31例早期乙肝肝纤维化患者的血清蛋白质谱,用biomarker Wizard软件筛选出早期肝纤维化的差异表达蛋白质谱峰,用Biomarker Pattern软件对差异蛋白峰值进行线性分析并建立诊断模型。结果在分子量为2000~20000Da的区间内检测出97个峰值,15个为差异蛋白峰,其中早期肝纤维化患者有3个上调,12个下调。用质荷比为M4799和M6125的差异蛋白联合建立的诊断模型最优,其特异度为80%(16/20),灵敏度为87%(27/31)。结论 SELDI-TOF-MS在筛选早期肝纤维化血清学标志物方面具有较高的灵敏度和特异性,作为肝纤维化早期筛选的方法尚需大宗病例证实及临床后续验证。  相似文献   

16.
目的:比较结直肠癌与正常人血清蛋白质谱的变化,筛选特异性蛋白标志物,建立结直肠癌诊断分类树模型.方法:收集血清样本133例(其中结直肠癌67例,正常人66例),随机分为建模组和验证组.运用弱阳离子纳米磁珠(magnetic bead-weak cation exchange,MB-WCX)联合基质辅助激光解吸离子飞行质谱(matrix-assistedlaser desorption/ionization time-of-flight massspectrometry,MALDI-TOF-MS),建立结直肠癌与正常人血清蛋白质谱.用Flex Analysis2.4软件收集数据,应用ClinproTools2.2软件对建模组34例结直肠癌和33例正常人血清差异蛋白质谱进行定量分析,应用Genetic Algorithm算法建立结直肠癌诊断模型,应用所获取的诊断模型对验证组样本(33例结直肠癌和33例正常人)进行分类诊断,以评价诊断模型的诊断价值.结果:通过比较分析结直肠癌与正常人血清蛋白质谱,发现共有33个差异蛋白峰(P<0.05),其中在结直肠癌中表达上调25个,表达下调8个.利用其中5个差异峰(Mr分别为759,3316,4645,4248,2645Dr)建立诊断模型,获得了94.12%(32/34)敏感性和96.97%(32/33)的特异性,经独立样本双盲验证,其灵敏度为93.94%(31/33),特异度为96.97%(32/33).结论:基于磁珠分离和MALDI-TOF-MS技术能直接检测出结直肠癌患者血清差异表达蛋白,建立的诊断模型具有较高的敏感性和特异性,对提高结直肠癌的诊断具有一定的临床意义.  相似文献   

17.
Endostatin is an endogenous inhibitor of angiogenesis and tumor growth in mice, which may be generated by proteolytic cleavage of collagen XVIII. In normal tissues, 2 variants of the endostatin precursor, namely the SHORT and LONG forms, regulate tissue specificity. We analyzed 53 human liver biopsies (18 hepatocellular carcinomas, 16 metastases of colorectal cancer, 3 cholangiocarcinomas, and 16 controls) by RNA dot blots, double-labeling immunohistochemistry, and in situ hybridization, using common and variant-specific probes. Tumor hepatocytes expressed the LONG form, whereas cholangiocarcinoma cells expressed the SHORT form, which was deposited in tumor basement membranes. Metastatic colorectal carcinoma cells did not express collagen XVIII. In the stromal compartment of primary and metastatic cancers, myofibroblasts and vascular endothelial cells expressed the SHORT form. Both basement membrane components, collagen IV and the SHORT collagen XVIII form, were codistributed and their mRNA levels strongly correlated (R =.75, P <.001). In addition, freshly isolated human hepatocytes expressed the LONG form and culture-activated stellate cells the SHORT form. Moreover, the full-length LONG form is a plasma protein. Thus, the LONG form is a hepatocyte-specific variant, and the SHORT form is a major component of the tumor extracellular matrix in primary and metastatic liver cancers. In the clinical context, the global expression of the endogenous endostatin precursor, collagen XVIII, in liver cancer results from the combined expression profiles of tumor cells, stromal cells, and nontumor hepatocytes at the advancing edge of the tumor, particular to each type of cancer.  相似文献   

18.
目的:探索大肠腺瘤与大肠腺癌差异表达的肿瘤相关基因.方法:利用基因芯片技术筛选大肠腺瘤与大肠腺癌差异表达的肿瘤相关基因,比较两组基因特点,寻找大肠腺癌重要致病基因,并以RT-PCR对部分差异表达基因进行检测来验证芯片结果.结果:(1)大肠腺瘤差异表达的肿瘤相关基因9个,均上调;(2)大肠腺癌差异表达的肿瘤相关基因47个...  相似文献   

19.
We present a case of multiple colorectal liver metastases with macroscopic portal vein thrombi. A 55-year-old woman presented to us with rectosigmoid cancer and presented with two liver metastases. The tumor in the posterior sector was associated with invasion of first order branches of the portal vein. We performed low anterior resection, hepatic posterior sectorectomy and partial left anterior sectorectomy. Both the colorectal cancer and liver tumors exhibited histological characteristics of moderately differentiated adenocarcinoma with a substantial amount of mucin production. The liver metastases were associated with prominent tumor thrombi in many branches of the portal vein. Stronger staining for endoglin (CD 105) than for Fas ligand (Fas L) and matrix metalloproteinase (MMP-2) was observed in both the colorectal cancer and metastatic liver tumor cells. Expression of the vascular endothelial growth factor within the tumor cells was seen in both the colorectal cancer as well as the metastatic liver tumor cells. Six months after the operation, she was diagnosed to have multiple, more than about 20 liver metastases, and in 9 months after the operation, the patient died. The colorectal cancer with liver metastases associated with portal vein tumor thrombosis was poor prognosis, found neoplastic microvessel formation.  相似文献   

20.
体外培养的肝癌细胞株与正常肝细胞株蛋白质的差异表达   总被引:5,自引:2,他引:5  
目的:运用SELDI蛋白质芯片技术分析体外培养的肝癌细胞株(HepG2)与正常肝细胞株(L02)蛋白质表达差异.方法:在体外培养HepG2和L02细胞株,收获细胞,将细胞用细胞裂解液裂解后,采用SELDI蛋白质芯片技术用IMAC3 及WCX2芯片检测HepG2、L02的蛋白质谱.结果:体外培养的肝癌细胞株与正常肝细胞株的蛋白质存在差异表达,IMAC3芯片共捕获61个蛋白,发现差异峰7个,与 L02细胞相比,其中3个差异蛋白在肝癌细胞中高表达,4个差异蛋白在肝癌细胞中低表达.WCX2芯片共捕获91个蛋白, 发现差异峰14个,其中3个差异蛋白在肝癌细胞中高表达,11 个差异蛋白在肝癌细胞中低表达.结论:SELDI蛋白芯片技术检测肝癌细胞株与正常肝细胞株蛋白质的差异表达方法简便,敏感性高,重复性好.  相似文献   

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