肺岩宁方调控MALAT1抑制A549肺癌细胞侵袭的机制研究

Mechanism study on Feiyanning Recipe inhibiting invasion of A549 lung cancer cells by regulating MALAT1

目的探讨肺岩宁方调控肺腺癌转移相关转录本1(MALAT1)抑制A549肺癌细胞侵袭的机制。方法向A549细胞转染LV-vector、LV-over/MALAT1慢病毒载体,应用肺岩宁方(0～500μg/ml)干预细胞,应用实时定量聚合酶链式反应(qRT-PCR)法检测MALAT1表达情况;应用四甲基偶氮唑盐(MTT)比色法检测肺岩宁方对肿瘤细胞增殖的影响;细胞侵袭实验检测肿瘤侵袭能力;Western blot法检测Wnt/β-catenin/EMT信号通路相关蛋白的表达。结果细胞侵袭实验结果显示,过表达MALAT1可增强A549细胞的侵袭能力,与正常对照组和慢病毒载体组相比,差异有统计学意义(P<0.01);与正常对照组相比,肺岩宁方可抑制A549细胞侵袭,差异有统计学意义(P<0.01),但过表达MALAT1后可减弱肺岩宁抑制细胞侵袭的能力。MTT结果显示,肺岩宁方可抑制肺癌A549细胞增殖,且呈浓度依赖性。qRT-PCR结果显示,肺岩宁方可下调MALAT1表达。Western blot结果显示,MALAT1可下调E-cadherin蛋白,上调N-cadherin、Snail及细胞核中β-catenin的蛋白水平,肺岩宁方能上调E-cadherin蛋白,下调N-cadherin、Snail及细胞核中β-catenin的蛋白水平,但过表达MALAT1可削弱肺岩宁对这些蛋白的调控作用。结论 MALAT1过表达通过激活Wnt/β-catenin通路,加速EMT进程促进A549肺癌细胞侵袭;肺岩宁方能够通过下调MALAT1,抑制Wnt/β-catenin/EMT信号轴,从而阻止A549肺癌细胞侵袭。

Objective To explore the mechanism of action of Feiyanning Recipe(FYN) in inhibiting invasion of the A549 lung cancer cells by regulating metastasis-associated lung adenocarcinoma transcript 1(MALAT1). Methods The A549 cells transfected with LV-vector and LV-over/MALAT1 were treated with FYN(0~500 μg/ml). The expression of MALAT1 was detected by qRT-PCR;the effect of FYN on tumor cell proliferation was determined by MTT colorimetric assay;the invasive ability of tumor was determined by cell invasion test;and the expression of Wnt/β-catenin/EMT signaling pathway related proteins in A549 cells was detected by Western blotting. Results Results of the cell invasion test indicated that MALAT1 overexpression strengthened the invasive ability of the A549 cells, and there was significant difference compared to the normal control group and the lentivirus vector group(P<0.01). Compared to the normal control group, FYN inhibited invasion of the A549 cells, with significant difference(P<0.01),but the ability of FYN to inhibit cell invasion was reduced after MALAT1 overexpression. Results of MTT colorimetric assay showed that FYN inhibited proliferation of the A549 cells in a concentration-dependent manner. Results of qRT-PCR indicated that FYN down-regulated the expression of MALAT1. Results of Western blotting showed that MALAT1 down-regulated E-cadherin and up-regulated N-cadherin,Snail and β-catenin in the nucleus,whereas FYN up-regulated E-cadherin and down-regulated N-cadherin,Snail and β-catenin in the nucleus,but MALAT1 overexpression reduced the regulatory effect of FYN on these proteins. Conclusion MALAT1 overexpression promotes lung cancer cell invasion by activating the Wnt/β-catenin pathway and accelerating the EMT progress. Feiyanning Recipe inhibits invasion of the A549 lung cancer cells by down-regulating MALAT1 and suppressing the Wnt/β-catenin/EMT signal axis.