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目的:探讨磷脂酰肌醇蛋白聚糖-5(glypican 5,GPC5)基因对人肺腺癌A549肿瘤起始细胞(cancer-initiating cells,CICs)生物学功能的影响。方法 :采用前期构建的过表达GPC5慢病毒载体稳定转染的细胞株GPC5-A549,无血清培养方法富集A549细胞中的CICs微球,通过定量分析检测微球大小,流式细胞术检测CD133阳性细胞比率,Matrigel和Tanswell小室检测细胞侵袭迁移能力,Western blot检测上皮细胞间质化相关蛋白表达。结果:与对照组相比,过表达GPC5后A549细胞中CICs微球形成数目明显减少,体积变小,CD133阳性率明显下降(P<0.05);细胞侵袭、迁移能力显著降低(P<0.05);E-cadherin的蛋白表达明显上调,Vimentin、N-cadherin的表达显著被抑制(P<0.05)。结论:GPC5过表达能够抑制肺腺癌细胞株A549中CICs的发生、侵袭和迁移,调控上皮细胞间质化相关蛋白表达。
Objective: To investigate the effect of glypican 5 (GPC5) gene on the biological function of human lung adenocarcinoma A549 tumor-initiating cells (CICs). METHODS: The cell line GPC5-A549 stably transfected with GPC5 overexpressing lentiviral vectors constructed in the previous period was used to enrich CICs microspheres in A549 cells by serum-free culture method. The size of microspheres was detected by quantitative analysis, and CD133 was detected by flow cytometry. The ratio of positive cells, Matrigel and Tanswell chambers were used to detect the invasion and migration of cells, and Western blot was used to detect the expression of epithelial-mesenchymal proteins. RESULTS: Compared with the control group, the number of CICs microspheres in A549 cells was significantly reduced after the GPC5 was overexpressed. The volume of the CICs microspheres was decreased and the CD133 positive rate was significantly decreased (P<0.05). The invasion and migration of cells were significantly decreased (P<0.05). The protein expression of E-cadherin was significantly up-regulated, and the expression of Vimentin and N-cadherin was significantly inhibited (P<0.05). Conclusion: Over-expression of GPC5 can inhibit the occurrence, invasion and migration of CICs in lung adenocarcinoma cell line A549 and regulate the expression of epithelial-mesenchymal proteins.