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胃癌中比较蛋白质组学的技术评估及分化相关蛋白的筛选

Technique appraisement of comparative proteomics and screening of differentiation-related protein in gastric carcinoma.

作者信息

Liu Yu, Li Yong, Tan Bi-bo, Zhao Qun, Fan Li-qiao, Zhang Zhi-dong, Li Zhao-xing

机构信息

Department of General Surgery, The Fourth Affiliated Hospital, Hebei Medical University, Shijiazhuang, China.

出版信息

Hepatogastroenterology. 2013 May;60(123):633-7. doi: 10.5754/hge12794.

Abstract

BACKGROUND/AIMS: Different differentiations of cancer have resulted in its unique biological characteristics. We screen and appraise differentially expressed proteins in different differentiated gastric adenocarcinoma with comparative proteomics technology in order to find regulatory factors of tumor differentiation related and finally reach the purpose of tumor differentiation reversal.

METHODOLOGY

With two-dimensional fluorescence difference gel electrophoresis (2-D DIGE) and liquid chromatography in conjunction with tandem mass spectrometry (LC–MS/MS), the differentially expressed proteins from 8 patients with different differentiated gastric adenocarcinoma were identified and some factors identified were verified with application of QPCR and Western blot techniques.

RESULTS

Significant differences in 35 protein spots were found and 48 kinds of proteins were identified. Other than structural proteins and non-specific protein, six possible proteins associated with tumor differentiation were determined - the serine protease inhibitor B1 (serine protease inhibitor, clade B, member 1, SERPINB1), calcium-phospholipid binding protein III (annexin A3), transcription factor Nm23-H1, adenine phosphoribosyl-transferase enzyme APRT (Adenine Phosphoribosyltransferase in APO and AMP), glutathione S-transferase P1-1 (GST-π-1), antimicrobial peptides Dermcidin-lL. The identified SERPINB1, annexin A3, Nm23-H1 and APRT were verified, confirming the expression of these factors was in line with proteomics identification.

CONCLUSIONS

Protein expression in different differentiated gastric adenocarcinoma was varied.

摘要

背景/目的:癌症的不同分化导致其具有独特的生物学特性。我们运用比较蛋白质组学技术筛选并评估不同分化程度的胃腺癌中差异表达的蛋白质,以寻找肿瘤分化相关的调控因子,最终达到逆转肿瘤分化的目的。

方法

采用二维荧光差异凝胶电泳(2-D DIGE)和液相色谱串联质谱(LC-MS/MS)技术,鉴定8例不同分化程度胃腺癌患者的差异表达蛋白质,并运用QPCR和蛋白质免疫印迹技术对部分鉴定出的因子进行验证。

结果

发现35个蛋白点存在显著差异,鉴定出48种蛋白质。除结构蛋白和非特异性蛋白外,确定了6种可能与肿瘤分化相关的蛋白质——丝氨酸蛋白酶抑制剂B1(丝氨酸蛋白酶抑制剂,B族,成员1,SERPINB1)、钙磷脂结合蛋白III(膜联蛋白A3)、转录因子Nm23-H1、腺嘌呤磷酸核糖转移酶APRT(载脂蛋白和AMP中的腺嘌呤磷酸核糖转移酶)、谷胱甘肽S-转移酶P1-1(GST-π-1)、抗菌肽Dermcidin-lL。对鉴定出的SERPINB1、膜联蛋白A3、Nm23-H1和APRT进行了验证,证实这些因子的表达与蛋白质组学鉴定结果一致。

结论

不同分化程度的胃腺癌中蛋白质表达存在差异。

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