摘要
目的探讨miR-195-5p及血管内皮生长因子(Vascular endothelial growthfactor,VEGF)在肺腺癌患者血清中的表达及miR-195-5p对肺腺癌细胞A549迁移能力和血管生成的影响。方法采用qRT-PCR法及ELISA法检测30例确诊肺腺癌患者及30例正常人血清miR-195-5p、磷脂酸磷酸酶2B(Phosphatidic acid phosphatase type 2B,PPAP2B)基因及VEGF蛋白的表达,通过脂质体Lipo-fectamine 3000转染miR-195-5p mimics至人肺泡腺癌基底上皮细胞(A549细胞),ELISA法检测control组和miR-195-5p mimics组细胞培养上清液中VEGF蛋白的表达,qRT-PCR法分析两组细胞miR-195-5p与PPAP2B基因的表达,Western blot检测两组细胞PPAP2B、VEGF蛋白的水平,Transwell实验分析miR-195-5p对A549细胞迁移能力的影响。结果与正常人比较,miR-195-5p在肺腺癌患者血清中低表达,而PPAP2B基因和VEGF蛋白在肺腺癌患者血清中高表达,其差异均有统计学意义(P<0.05)。高表达miR-195-5p后A549细胞培养上清液中的VEGF蛋白表达下调,细胞PPAP2B基因表达降低,细胞迁移能力明显降低。结论miR-195-5p降低VEGF蛋白的表达及抑制A549细胞的迁移,其作用机制可能与调控靶基因PPAP2B有关,两者在肺腺癌病理过程中可能发挥重要作用。
Objective To investigate the expressions of miR-195-5p and vascular endothelial growth factor(VEGF)in serum of patients with lung adenocarcinoma and the effects of miR-195-5p on the migration and angiogenesis of A549 cells.Methods The expressions of miR-195-5p and phosphatidyl acid phosphatase type 2B(PPAP2B)gene and VEGF protein level in serum of 30 patients with pulmonary adenocarcinoma and 30 normal individuals were detected by qRT-PCR and ELISA.MiR-195-5p were transfected by lipo-fectamine 3000 to human alveolar adenocarcinoma basal epithelium(A549 cells).ELISA was used to detect the expression of VEGF protein in the cells culture supernatant of the control group and miR-195-5p mimics group,and qRT-PCR was used to analyze miR-195-5p and PPAP2B gene expressions.Western blot was used to detect the expression of PPAP2B and VEGF proteins in the two groups of cells.Transwell migrationassay was used to analyze the effect of miR-195-5p on the migration capacity of A549 cells.Results In the serum of patients with lung adenocarcinoma,the expression of miR-195-5p was lower,while PPAP2B gene and VEGF protein were highly expressed.The difference were all statistically significant(P<0.05).After overexpression of miR-195-5p,the expression of VEGF protein in A549 cells culture supernatant were down-regulated,PPAP2B expression was also reduced in cells,and the cells migration ability were significantly reduced.Conclusion MiR-195-5p can reduce the expression of VEGF and inhibit the migration of A549 cells.Its mechanism may be related to the regulation of PPAP2B,which may play an important role in the process of lung adenocarcinoma.
作者
李才弘
杨华军
刘长路
刘娅
黄婷
龚玲
佘安俊
刘代顺
Li Caihong;Yang Huajun;Liu Changlu;Liu Ya;Huang Ting;Gong Ling;She Anjun;Liu Daishun(Department of Pulmonary and Critical Care Medicine,The Third Affiliated Hospital of Zunyi Medical University,Zunyi Guizhou 563002,China;Zunyi Institute of Respiratory Diseases,Zunyi Guizhou 563002,China)
出处
《遵义医科大学学报》
2020年第1期64-68,共5页
Journal of Zunyi Medical University
基金
遵义市科技局基金资助项目(NO:遵市科合社字[2018]165
NO:遵市科合[2016]39
NO:遵市科合[2015]17)。
