摘要
目的 探讨整合素连接激酶(ILK)促进肺癌细胞侵袭的相关分子机制.方法 通过细胞转染、siRNA干扰、细胞划痕实验、实时定量PCR、Western Blot方法探讨ILK和基质金属蛋白酶9(MMP-9)在肺癌A549细胞中的表达及相互关系.结果 在肺癌A549细胞系中,过度表达的ILK诱导MMP-9的表达(P<0.01);加入MMP-9抑制剂多西环素显著影响了转染组细胞划痕愈合能力(P<0.01),加入抗-MMP-9中和抗体则严重阻碍了细胞迁移能力(P<0.01),体外基底膜侵袭实验亦得到了相同的结果(P<0.01).ILK的过度表达促进磷酸化和核因子-κB(NF-κB)亚单位p65的核易位(P<0.01),并且NF-κB抑制剂BAY11-7028和NF-κBp65siRNA能抑制ILK高表达细胞系中MMP-9的上调(P<0.01).结论 本研究结果表明,ILK的过度表达可促进肺癌细胞迁移和侵袭,并且是通过NF-κB途径介导MMP-9上调来实现这一过程.
Objective To investigate the role of integrin-linked kinase (ILK) on migration and invasion of lung cancer cell by upregulating matrix metalloproteinase-9 through nuclear factor-κB (NF-κB) pathway.Methods A549 cell line were overexpressed ILK and matrix metalloproteinase-9 (MMP-9) confirmed by cell transfection,siRNA interference,cell scratch test,real-time quantitative PCR and Western Blot.Results Over-expression of ILK stimulated MMP-9 expression in lung cancer cells(P < 0.01).The addition of MMP-9 inhibitor doxycycline and anti-MMP-9 neutralizing antibody significantly impaired the wound healing capacity of ILK-transfected cells(P < 0.01),as well as by in vitro matrigel invasion assay (P < 0.01).In addition,overexpression ILK induced phosphorylation and nuclear translocation of nuclear factor-κB subunit p65.Upregulation of MMP-9 was severely abolished by either BAY 11-7028,a specific NF-κB inhibitor,or siRNA targeted to NF-κB p65 in ILK over-expression cells.Conclusion The finding indicate that over-expression of ILK can promote the migration and invasion of lung cancer cell,and upregulate MMP-9 through the NF-κB pathway.
出处
《中国综合临床》
2014年第3期243-248,共6页
Clinical Medicine of China
基金
辽宁省科学技术计划项目(2012225016)
辽宁省自然科学基金(20092097)