RNA干扰阻断CXCR4基因对胰腺癌AsPC-1细胞侵袭转移的影响  

Effect of RNA inference mediated CXCR4 knockdown on metastasis of As PC-1 cells

作　　者：黄继英[1] 高振军[1] 沈曼茹[1] 孟鑫[2] 

机构地区：[1]复旦大学附属中山医院青浦分院消化科,上海市201700 [2]复旦大学附属中山医院青浦分院医院感染管理科,上海市201700

出　　处：《世界华人消化杂志》2016年第9期1342-1348,共7页World Chinese Journal of Digestology

基　　金：上海市卫生和计划生育委员会基金资助项目;No.201440607;上海市青浦区卫生和计划生育委员会基金资助项目;No.W2014-01~~

摘　　要：目的:观察针对CXCR4基因慢病毒Lentiviral(LV)介导的RNAi转染胰腺癌细胞株As PC-1后细胞增殖、迁移、侵袭的影响,探讨可能机制.方法:人CXCR4基因干扰靶点设计后成功转染人胰腺癌细胞系As PC-1细胞,然后分为转染组(LV-si CXCR4-1)、阴性对照组(As PC-1-LV-con)和空白对照组(AsP C-1细胞),实验组加入基质细胞衍生因子-1α(stromal cell-derived factor-1α,SDF-1α),MTT法检测细胞增殖,迁移实验检测细胞迁移,Transwell检测细胞的体外侵袭,Western blot检测血管内皮生长因子(vascular endothelial growth factor,VEGF)、基质蛋白金属酶(matrix metalloproteinases,MMP)2和MMP9的表达.结果:在SFM或10%NCS培养条件下,SDF-1α均可刺激As PC-1细胞、As PC-1-LV-Con细胞增殖(P<0.01),对LV-si CXCR4-1增殖均无明显促进作用;SDF-1α可促进As PC-1、As PC-1-LV-C o n细胞迁移和侵袭(P<0.01),对LVsi CXCR4-1无明显促进作用;在SDF-1α作用下,LV-si CXCR4-1细胞的MMP9及VEGF的蛋白表达低于As PC-1、As PC-1-LV-Con细胞(P<0.01).结论:CXCR4基因RNAi可显著抑制SDF-1α引起的As PC-1细胞的增殖、体外迁移和侵袭,进一步证实SDF-1/CXCR4受体配体系统参与了胰腺癌的增殖、侵袭及转移,通过RNAi技术可抑制胰腺癌细胞的增殖、侵袭及转移.AIM： To explore the changes in proliferation, migration, and invasion of pancreatic cancer line AsPC-1 after CXCR4 is knocked down by RNA interference, and to validate the potential mechanism underlying these changes.METHODS： AsPC-1 cells were transfected with recombinant lentivirus containing specific shRNA targeting CXCR-4 and included in an experimental group（LV-siCXCR4-1）, whereas AsPC-1 cells treated with recombinant lentivirus containing scramble shRNA were used as a negative control（AsPC-1-LV-con）. Besides, non-treated AsPC-1 cells were used as a blank control（AsPC-1）. After cell cultures were treated with SDF-1α, cell proliferation, migration and invasion in vitro were analyzed, and expression of vascular endothelial growth factor（VEGF）, matrix metalloproteinases（MMP）-2 and-9 were quantified.RESULTS： Under the culture conditions with SFM or 10% normal calf serum, SDF-1α could stimulate the proliferation of AsPC-1 cells and AsPC-1-LV-Con cells（P 0.01）, but had no significant effect on the proliferation of AsPC-1-LV-siCXCR4-1 cells. SDF-1α displayed a promoting effect on the migration and invasion of AsPC-1 and AsPC-1-LV-Con cells（P 0.01）, but showed no significant effect in LV-siCXCR4-1 cells. In the presence of SDF-1α, the protein expression of MMP9 and VEGF in LV-siCXCR4-1 cells was less than that in AsPC-1-LV-Con cells（P 0.01）.CONCLUSION： RNA interference targeting CXCR-4 could significantly suppress excessive proliferation, migration and invasion of AsPC-1 cells induced by SDF-1α intervention, suggesting that CXCR4/SDF-1 axis contributes to proliferation, invasion and metastasis of pancreatic cancer. RNA interference could be used as a promising approach for therapy of pancreatic cancer.

关 键 词：胰腺肿瘤 基质细胞衍生因子-1 受体 CXCR4 RNA干扰 

分 类 号：R735.9[医药卫生—肿瘤]