[1]李勇,王力利,康爱文,等.应用蛋白质组学技术筛选鉴定锌指蛋白139调控的胃癌转移相关蛋白质[J].南方医科大学学报,2015,(03):360.
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应用蛋白质组学技术筛选鉴定锌指蛋白139调控的胃癌转移相关蛋白质()
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《南方医科大学学报》[ISSN:1673-4254/CN:44-1627/R]

卷:
期数:
2015年03期
页码:
360
栏目:
出版日期:
2015-03-20

文章信息/Info

Title:
Screening and identification of proteins related to gastric cancer metastasis with
comparative proteomics
作者:
李勇王力利康爱文范立侨赵群檀碧波郝英杰刘庆伟
关键词:
胃癌锌指蛋白139原位移植裸鼠淋巴结转移蛋白质组学
Keywords:
gastric cancer zinc finger protein 139 orthotopic transplantation model of nude mice lymph node metastasesproteomics
摘要:
目的以胃癌SGC7901细胞原位移裸鼠模型为对象,通过荧光双向差异凝胶电泳(2D-DIGE)结合液质联用(LC-MS)质谱
分析技术鉴定ZNF139 调控的胃癌转移相关蛋白质。方法合成针对ZNF139 的小干扰RNA(ZNF139-siRNA),以
ZNF139-siRNA转染人胃癌细胞株SGC7901,G418筛选。以ZNF139-siRNA质粒转染的胃癌细胞、阴性质粒转染的胃癌细胞
及普通胃癌细胞分别进行裸鼠胃癌原位移植。造模成功后取出原位移植瘤及腹腔转移淋巴结。荧光双向差异凝胶电泳
(2D-DIGE)技术分别分离ZNF139-siRNA 各组原位移植瘤及腹腔转移淋巴结蛋白质;选定差异点,胶内酶解后,液质联用
(LC-MS)质谱分析技术鉴定蛋白质。蛋白印迹(Western blot)技术验证差异蛋白质的表达。结果转染ZNF139-siRNA质粒后
SGC7901细胞中ZNF139的表达受到有效抑制。与阴性质粒组及空白组比较,阳性质粒组原位移植瘤生长更慢(P<0.05),且腹
腔淋巴结转移率更低(P<0.05)。蛋白质组学结果发现在阳性质粒组原发灶中Fascin、hnRNPA2/B1表达下调,ANXA1表达上
调;阳性质粒组转移淋巴结中ANXA5表达下调(P<0.05)。Western blot验证结果与蛋白质组学结果相符。结论ZNF139可能
通过调节Fascin、hnRNPA2/B1、ANXA1、ANXA5促进胃癌淋巴结转移。
Abstract:
Objective To screen and identify the proteins related with tumor metastasis of gastric cancer in a nude mouse model
bearing orthotopic transplanted tumor. Methods Zinc finger protein 139 (ZNF139)-specific siRNA was synthesized and
transfected into gastric cancer cell line SGC7901, which was then screened by G418. ZNF139-siRNA-transfected cells, negative
plasmid-transfected cells and untreated SGC7901 cells were orthotopically transplanted separately on the stomach wall of
BALB/c nude mice. The primary tumors and metastatic lymph nodes were harvested to separate the proteins by 2-D
fluorescence difference gel electrophoresis (2-D DIGE); after gel digestion, the differential proteins were subjected to liquid
chromatography-mass spectrometry (LC-MS) for identification and their functions were analyzed. Western blotting was
performed to verify the identified proteins. Results ZNF139 expression was effectively inhibited in siRNA-transfected
SGC7901 cells. ZNF139-siRNA-transfected cells showed obviously suppressed tumor growth with a lowered lymph node
metastasis rate in nude mice compared with untreated cells and the negative control cells (P<0.05). Proteomic study with 2-D
DIGE showed that fascin and hnRNPA2/B1 were down-regulated while ANXA1 was up-regulated in the primary tumors, and
ANXA5 was down-regulated in the metastatic lymph nodes in ZNF139-siRNA-transfected group. Western blotting confirmed
the results of proteomic analysis. Conclusion ZNF139 gene may promote lymph node metastasis of gastric cancer by
regulating fascin, hnRNPA2/B1, ANXA1, and ANXA5.

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更新日期/Last Update: 1900-01-01