沉默NGAL基因对肺癌细胞迁移及侵袭的影响  

Effect of Inhibiting NGAL Gene Expression on A549 Lung Cancer Cell Migration and Invasion

作　　者：唐健[1,2] 李捷[3] 李少军[2] 李璟波[2] 于长海[2] 尉承泽[1] 

机构地区：[1]军事医学科学院附属医院,北京100071 [2]解放军总医院第一附属医院胸心外科,北京100048 [3]解放军总医院胸外科,北京100853

出　　处：《中国肺癌杂志》2015年第4期187-192,共6页Chinese Journal of Lung Cancer

摘　　要：背景与目的检测NGAL在肺癌组织的表达,观察小干扰RNA(si RNA)沉默NGAL基因表达对肺癌细胞增殖及侵袭能力的影响。方法应用免疫组织化学检测肺癌组织中NGAL表达。通过RNAi技术干扰NGAL后,运用定量PCR、Western blot技术,观察干扰NGAL基因效率。干扰NGAL后,MTT检测细胞增殖、Transwell实验检测细胞迁移侵袭能力,细胞划痕愈合能力,免疫荧光及Western blot方法检测上皮细胞-间质细胞转化(epithelial-mesenchymal transition,EMT)相关的蛋白E-cadherin及Vimentin的表达情况。结果肺癌中NGAL阳性率高于癌旁组织(P<0.01)。NGAL-si RNA转染组,癌细胞中NGAL基因及蛋白水平表达都受到明显的抑制。NGAL-si RNA转染组细胞增殖能力,迁移及侵袭能力相对于对照组都降低,差异具有统计学意义(P<0.05)。干扰NGAL,E-cadherin的表达升高,Vimentin的表达水平降低。干扰NGAL,MMP-2和MMP-9蛋白表达水平降低。结论 NGAL在肺癌组织高表达,且在肺癌细胞的增殖、侵袭和迁移及EMT过程中起着重要作用。NGAL可能是肺癌浸润和转移的一个重要参考指标,可为肺癌治疗提供潜在的靶点。Background and objective To detect the expression of neutrophil gelatinase-assoeiated lipocalin （NGAL） in the different differentiations of lung cancer tissues and to study the mechanism of invasion ofA549 cells affected by NGAL. Methods The expression of NGAL was detected by immunochemistry in lung cancer tissue and the tissue around edge of the cancer. The effect of NGAL expression on A549 cells was observed by using qRT-PCR and Western blot. The abilities of invasion and metastasis were evaluated by transwell invasion and migration assay, and cell scratch assay in vitro. The protein expression of E-cadherin, Vimentin was measured by immunofluoresence and Western blot. Results The positive expression rate of NGAL was 76.32% （58/76） in the lung cancer, 13.3% （4/30） in adjacent tissue by immunochemistry. NGAL expres- sion levels in the lung cancer tissues were significantly higher than that in adjacent tissues. The rate of migration and invasion in NGAL-siRNA group was 60.4%＋6.4% compared to 50.5%＋4.4% in the control group, there was a significant difference （P〈0.05）. Vimentin was suppressed, and E-cadherin was upregulated when NGAL was inhibited. MMP-2 and MMP-9 de- creased when NGAL was knocked down. Conclusion The expression level of NGAL is highly expressed in lung cancer. NGAL may be one of important indicators involved in lung cancer infiltrated and transferred. NGAL might be one of potential targets for lung cancer treatment.

关 键 词：肺肿瘤 NGAL基因 迁移 侵袭 上皮细胞-间质细胞转化 

分 类 号：R734.2[医药卫生—肿瘤]