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

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。

Screening and identification of proteins related to gastric cancer metastasis withcomparative proteomics

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.