小干扰RNA抑制真核翻译起始因子5A2对MKN28胃癌细胞恶性生物学行为的影响  

Effects of Eukaryotic Translation Initiation Factor 5A2 Down-regulation by Small Interfering RNA on Aggressiveness of MKN28 Human Gastric Cancer Cell

作　　者：孟庆彬[1,2] 于健春[1] 康维明[1] 马志强[1] 周立[1] 叶欣[1] 曹战江[1] 田树波[1] 

机构地区：[1]中国医学科学院,北京协和医学院,北京协和医院基本外科,北京100730 [2]武汉市第一医院胃肠外科,武汉430022

出　　处：《中国医学科学院学报》2014年第5期482-487,共6页Acta Academiae Medicinae Sinicae

基　　金：北京自然科学基金(7132209);北京协和医学院研究生创新基金(2011-1002-017)~~

摘　　要：目的研究真核翻译起始因子5A2(EIF5A2)在胃癌细胞增殖及迁移侵袭调控中的作用及其可能机制。方法采用实时荧光定量PCR(qRT-PCR)及Western blot法检测EIF5A2在MKN28、HGC27细胞及正常胃黏膜上皮细胞(GES-1)内的表达,评估小干扰RNA(siRNA)对EIF5A2的抑制效果。检测EIF5A2蛋白在2例人胃癌及匹配的非癌胃黏膜组织的表达。采用细胞计数试剂盒(CCK-8)法检测细胞增殖,Transwell法检测细胞迁移及侵袭力,Western blot法检测CyclinD1、CyclinD3、基质金属蛋白酶-9(MMP-9)、E-cadherin、Vimintin、C-myc及转移相关蛋白1(MTA1)的表达水平。结果 EIF5A2在MKN28细胞及人胃癌组织内呈高表达。EIF5A2 siRNA#1可明显抑制MKN28细胞内EIF5A2 mRNA及蛋白的表达。抑制EIF5A2后可明显抑制MKN28细胞增殖(P均<0.01)、迁移(P<0.001)及侵袭能力(P<0.001)。抑制EIF5A2后,Vimentin表达下调,E-cadherin表达上调,CyclinD1、CyclinD3、MTA1及C-myc表达明显受抑制。结论siRNA下调EIF5A2可能通过抑制CyclinD1、CyclinD3抑制MKN28细胞增殖,并通过抑制MTA1、C-myc及上皮间质转化抑制MKN28细胞迁移及侵袭能力。Objective To investigate the effects of eukaryotic translation initiation factor 5A2 （EIFSA2） down-regulation by small interfering RNA （siRNA） on aggressiveness of human gastric cancer cell and its potential mechanisms. Methods The expressions of EIF5A2 in human gastric cancer cell lines （MKN28 and HGC27 ） and immortalized gastric mucosal epithelial cells （GES-1） were measured by real-time quantitative reverse tran- scription polymerase chain reaction （qRT-PCR） and Western blotting. EIFSA2 gene in MKN28 cells was si- lenced by RNA interference and the inhibitory effect was evaluated by both qRT-PCR and Western blotting. Cell proliferation was assessed by CCK-8 assay. Cell migration and invasion were assessed by Transwell assay. The possible downstream targets of EIF5A2, such as CyclinD1, CyclinD3, matrix metallopeptidase-9 （ MMP-9 ）, E-cadherin, vimintin, C-myc, and metastasis-associated protein 1 （MTA1） expression levels, were examined by Western blotting. Results High expressions of EIFSA2 were found in MKN28 cells and human gastric adeno- carcinoma tissues. Both EIF5A2 mRNA and protein expression in MKN28 cells were significantly down-regulated by siRNA#1 and siRNA#2, especially siRNA#1. Knockdown of EIFSA2 caused an apparent suppression of MKN28 cell proliferation （ all P 〈 0.01 ）, migration （ P 〈 0. 001 ）, and invasion （ P 〈 0. 001 ） . After the knockdown of EIFSA2 in MKN28 cells, E-cadherin levels were upregulated, whereas vimentin, Cyclin D1, Cyclin D3, C-myc and MTA1 levels were downregulated. Conclusion Knockdown of EIF5A2 may inhibit MKN28 cell proliferation by downregulating the CyclinD1 and CyclinD3 and suppressing the cell migration and in- vasion by inhibiting MTA1, C-myc and epithelial-mesenchymal transition.

关 键 词：真核翻译起始因子5A2 胃癌 小干扰RNA 增殖 转移 

分 类 号：R735.2[医药卫生—肿瘤]