CAJ | 학술논문

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.
목적 탐토정합소련접격매(ILK)촉진폐암세포침습적상관분자궤제.방법 통과세포전염、siRNA간우、세포화흔실험、실시정량PCR、Western Blot방법탐토ILK화기질금속단백매9(MMP-9)재폐암A549세포중적표체급상호관계.결과 재폐암A549세포계중,과도표체적ILK유도MMP-9적표체(P＜0.01)；가입MMP-9억제제다서배소현저영향료전염조세포화흔유합능력(P＜0.01),가입항-MMP-9중화항체칙엄중조애료세포천이능력(P＜0.01),체외기저막침습실험역득도료상동적결과(P＜0.01).ILK적과도표체촉진린산화화핵인자-κB(NF-κB)아단위p65적핵역위(P＜0.01),병차NF-κB억제제BAY11-7028화NF-κBp65siRNA능억제ILK고표체세포계중MMP-9적상조(P＜0.01).결론 본연구결과표명,ILK적과도표체가촉진폐암세포천이화침습,병차시통과NF-κB도경개도MMP-9상조래실현저일과정.
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.