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目的:探讨非小细胞肺癌患者血清中MMP-9表达与肿瘤转移的关系;MMP-9siRNA对肺腺癌细胞系A549中MMP-9表达及细胞侵袭与转移抑制作用的影响。方法:ELISA法检测32例非小细胞肺癌(NSCLC)患者及20名健康者空腹血清MMP-9浓度,同时设计合成针对MMP-9的特异性siRNA,采用oligofectamine转染A549细胞,RT-PCR法检测转染细胞后MMP-9mRNA的表达,Transwell实验检测细胞侵袭情况。结果:NSCLC患者血清中MMP-9浓度〔(63.44±7.84)ng/mL〕比正常对照组〔(21.57±3.29)ng/mL〕明显升高(P<0.01),且浓度随病理分期增加而增强〔Ⅰ/Ⅱ:(45.40±5.30)ng/mL,Ⅲ/Ⅳ:(89.80±5.02)ng/mL,P<0.01〕,发生淋巴结转移的患者血清MMP-9浓度〔(82.26±5.09)ng/mL〕明显高于未发生淋巴结转移〔(35.92±4.68)ng/mL〕的患者P<0.01;RT-PCR法显示,转染MMP-9siRNA后,细胞中MMP-9mRNA表达较阴性对照组和空白组明显降低,P<0.05。RNA干扰MMP-9基因后A549细胞的侵袭能力也有明显下降,P<0.01。结论:MMP-9可作为NSCLC患者有无发生浸润转移的良好预测指标,靶向MMP-9的siRNA不仅能特异性的降低MMP-9mRNA及蛋白的表达,且能抑制人A549细胞的侵袭和迁移。
Objective: To investigate the relationship between MMP-9 expression and tumor metastasis in patients with non-small cell lung cancer and the effect of MMP-9 siRNA on the expression of MMP-9 and cell invasion and metastasis in human lung adenocarcinoma cell line A549. Methods: The fasting serum MMP-9 levels in 32 NSCLC patients and 20 healthy volunteers were detected by ELISA. At the same time, specific siRNA targeting MMP-9 was designed and synthesized. A549 cells were transfected with oligofectamine. RT-PCR The expression of MMP-9 mRNA in transfected cells was detected, and the cell invasion was detected by Transwell assay. Results: Serum levels of MMP-9 in NSCLC patients were significantly higher than those in the control group [(63.44 ± 7.84) ng / mL 〔(21.57 ± 3.29) ng / mL〕 (P <0.01) The level of serum MMP-9 in patients with lymph node metastasis 〔(82.26 ± 5.09) ng〕 was significantly higher than that in patients with lymph node metastasis 〔(Ⅰ) / Ⅱ: (45.40 ± 5.30) ng / mL, / mL] were significantly higher than those without lymph node metastasis [(35.92 ± 4.68) ng / mL〕 P <0.01; RT-PCR showed that after transfected with MMP-9siRNA, MMP-9mRNA expression in cells compared with negative control group and The blank group was significantly lower, P <0.05. The invasion ability of A549 cells after RNA interference with MMP-9 gene was also significantly decreased (P <0.01). Conclusion: MMP-9 can be used as a good predictor of invasion and metastasis in patients with NSCLC. SiRNA targeting MMP-9 can not only specifically reduce the expression of MMP-9 mRNA and protein, but also inhibit the invasion and migration of human A549 cells .