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采用二维凝胶电泳和基质辅助激光解吸/电离飞行时间质谱法检测人肝癌组织中Raf激酶抑制蛋白的表达

Expression of Raf kinase inhibitor protein in human hepatoma tissues by two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization time-of-flight methods.

作者信息

Tsao D A, Shiau Y F, Tseng C S, Chang H R

机构信息

Department Medical Technology, Fooyin University, Kaohsiung City, Taiwan.

Department of Hepato-Gastroenterology, Center-of-Hepato-Gastroenterology, Yuan's General Hospital, Kaohsiung City, Taiwan.

出版信息

Indian J Cancer. 2016 Apr-Jun;53(2):216-219. doi: 10.4103/0019-509X.197730.

Abstract

PURPOSE

Hepatocellular carcinoma (HCC) is the most common malignant liver tumor. To reduce the mortality and improve the effectiveness of therapy, it is important to search for changes in tumor-specific biomarkers whose function may involve in disease progression and which may be useful as potential therapeutic targets. Materials and Mehtods: In this study, we use two-dimensional polyacrylamide gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry to observe proteome alterations of 12 tissue pairs isolated from HCC patients: Normal and tumorous tissue. Comparing the tissue types with each other, 40 protein spots corresponding to fifteen differentially expressed between normal and cancer part of HCC patients.

RESULTS

Raf kinase inhibitor protein (RKIP), an inhibitor of Raf-mediated activation of mitogen-activated protein kinase/extracellular signal-regulated kinase, may play an important role in cancer metastasis and cell proliferation and migration of human hepatoma cells. RKIP may be considered as a marker for HCC, because its expression level changes considerably in HCC compared with normal tissue. In addition, we used the methods of Western blotting and real time-polymerase chain reaction to analysis the protein expression and gene expression of RKIP. The result showed RKIP protein and gene expression in tumor part liver tissues of HCC patient is lower than peritumorous non-neoplastic liver tissue of the corresponding HCC samples.

CONCLUSION

These results strongly suggest that RKIP may be considered to be a marker for HCC and RKIP are down-regulated in liver cancer cell.

摘要

目的

肝细胞癌(HCC)是最常见的肝脏恶性肿瘤。为降低死亡率并提高治疗效果,寻找肿瘤特异性生物标志物的变化非常重要,这些标志物的功能可能涉及疾病进展,并且可能作为潜在的治疗靶点。材料与方法:在本研究中,我们使用二维聚丙烯酰胺凝胶电泳(2-DE)和基质辅助激光解吸/电离飞行时间质谱来观察从HCC患者分离的12对组织(正常组织和肿瘤组织)的蛋白质组变化。将这些组织类型相互比较,发现40个蛋白点对应于HCC患者正常组织和癌组织之间差异表达的15种蛋白质。

结果

Raf激酶抑制蛋白(RKIP)是Raf介导的丝裂原活化蛋白激酶/细胞外信号调节激酶激活的抑制剂,可能在癌症转移以及人肝癌细胞的增殖和迁移中起重要作用。RKIP可被视为HCC的标志物,因为与正常组织相比,其在HCC中的表达水平有显著变化。此外,我们使用蛋白质印迹法和实时聚合酶链反应方法分析RKIP的蛋白表达和基因表达。结果显示,HCC患者肿瘤部分肝组织中RKIP的蛋白和基因表达低于相应HCC样本的瘤旁非肿瘤性肝组织。

结论

这些结果强烈表明RKIP可被视为HCC的标志物,并且RKIP在肝癌细胞中表达下调。

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