中文摘要：

目的研究Wnt5a在A549细胞球增殖和EMT中的作用及相关机制。方法利用无血清培养出A549细胞球后应用流式细胞术对其进行CD133＋CD44＋表达检测。实验分为对照组、Wn5a组、p38组和Wnt5a＋p38组。应用Western blot观察各组Wnt5a、p-p38、β-catenin、Vimentin和E-cadherin蛋白的表达变化,并利用MTT和裸鼠成瘤实验观察Wnt5a shRNA慢病毒感染前后A549细胞球的生长抑制率和成瘤能力。结果流式细胞术检测A549细胞球中CD133＋CD44＋阳性率为60.5%;Wnt5a蛋白在Wnt5a组和Wnt5a＋p38组中的表达较对照组明显下调（P〈0.05）,p-p38蛋白、β-catenin蛋白、Vimentin蛋白在Wnt5a组、p38组和Wnt5a＋p38组的表达明显低于对照组（P〈0.05）,而E-cadherin蛋白在上述3组中的表达显著高于对照组（P〈0.05）。Wnt5a shRNA慢病毒感染A549细胞球12、24 h和48 h时抑制率显著高于对照组（P〈0.05）,并且该细胞的成瘤能力较对照组显著下调。结论下调Wnt5a表达可抑制A549细胞球的增殖及EMT能力,可望成为治疗非小细胞肺癌的靶点。

英文摘要：

Objective To deternine the role and mechanism of Wnt5 a regulating the proliferation and epithelial mesenchymal transition（EMT） in lung cancer A549 spheres. Methods A549 spheres were cultured in serum-free medium for 2 weeks. CD133＋CD44＋rate of those cells was detected by FACS. Four experimental groups including a control group,a Wnt5 a group,a p38 group and a Wnt5 a ＋ p38 group were designed. The protein expression of Wnt5 a,p-p38,β-catenin,vimentin and E-cadherin among the 4 groups was detected by Western blotting. The cell inhibitory rate and tumorigenicity were detected after Wnt5 a shRNA lentivirus transfection of A549 spheres. Results CD133＋CD44＋rate was 60. 5% in A549 spheres and 1. 65% in A549 cells. Wnt5 a protein expression was significantly lower in the Wnt5 a and Wnt5 a ＋ p38 groups than in the control group（P〈0. 05）. Compared to the control group,the protein expression of p-p38,β-catenin and vimentin was obviously lower but that of E-cadherin was higher in the Wnt5 a,p38 and Wnt5 a ＋ p38 groups（ P〈0. 05）. The cell inhibitory rate was significantly higher and tumor size in node mice was remarkably smaller at12,24 and 48 h after Wnt5 a shRNA lentivirus transfection（P〈0. 05）. Conclusion Wnt5 a expression downregulation inhibits the proliferation and EMT of A549 spheres,and Wnt5 a might be a novel target for treatment of non-small-cell lung cancer.