hsa-miR-195-5p靶向血管内皮生长因子A对人胰腺癌PANC-1细胞侵袭和迁移的调控作用  

hsa-miR-195-5p Regulates the Invasion and Migration of Human Pan- creatic Carcinoma PANC-1 Cells by Targeting Vascular Endothelial Growth Factor A

作　　者：曹静 周鹏飞[2] CAO Jing;ZHOU Pengfei(Department of Laboratory,Baoji City Medical Hospital,Baoji,Shaanxi,721000,China;Baoji Maternal and Child Health Hospital Baoji,Baoji,Shaanxi,721000,China)

机构地区：[1]宝鸡市中医医院检验科,陕西省宝鸡市721000 [2]宝鸡市妇幼保健院,陕西省宝鸡市721000

出　　处：《医学分子生物学杂志》2018年第5期278-282,共5页Journal of Medical Molecular Biology

基　　金：陕西省2014年卫生科研项目（No.2014D84）

摘　　要：目的探索hsa-miR-195-5p对人胰腺癌PANC-1细胞侵袭和迁移的影响。方法将miR-195mim-ic和pc．VEGFA过表达载体单独或同时转入PANC-1细胞，以提高miR-195水平和过表达血管内皮生长因子A（vascularendothelialgrowthfactorA，VEGFA）。PANC-1细胞分为4组：PANC-1（对照）组、miR，195mimic组、pc-VEGFA组和miR-195mimic’pc-VEGFA组。实时定量PCR（quantitativereal-timereversetran-scriptionPCR，qRT-PCR）检测miR-195水平和VEGFA的mRNA水平。荧光素酶实验验证miR-195与VEG-FA的靶向关系。蛋白印迹检测VEGFA蛋白水平。Transwell检测细胞侵袭，划痕实验分析细胞迁移。结果与对照组相比，miR-195mimic组miR-195水平显著升高（P〈0．001）。miR-195mimic组VEGFA的mR-NA水平显著低于对照组（P〈0．001）。荧光素酶实验显示hsa-miR-195．5p可直接靶向VEGFA。miR-195mimic可抑制pc-VEGFA对VEGFA表达的促进作用（P〈0．01）。miR-195mimic组细胞侵袭显著低于对照组（P〈0．01）。pc-VEGFA组细胞侵袭显著高于对照组（P〈0．01）。与pc-VEGFA组相比，miR-195mimic＋pc-VEGFA组细胞侵袭显著降低（P〈0．01）。与对照组相比，miR-195mimic组细胞迁移显著下降，pe-VEGFA组细胞迁移显著升高（P〈0．01）。miR-195mimic＋pc-VEGFA组细胞迁移显著低于pc-VEGFA组（P〈0．01）。结论hsa-miR-195-5p可通过靶向VEGFA减弱人胰腺癌PANC-1细胞侵袭和迁移。Objective The study aimed to explore the effect of hsa-miR-195-5 p on the invasion and migration of human pancreatic carcinoma PANC-1 cells. Methods MiR-195 mimic and pc-VEGFA was tranfected into PANC-1 cells to increase the levels of miR-195 and expression of vascular endothelial growth factor A （VEGFA） . PANC-I cells were divided into four groups： PANC- 1 （control） group, miR-195 mimic group, pc-VEGFA group and miR-195 mimic ＋ pc-VEGFA group. The level of miR-195 and mRNA level of VEGFA were detected by quantitative real-time re- verse transcription PCR （qRT-PCR） . The targeted relationship of miR-195 and VEGFA was verified by luciferase assay. The protein level of VEGFA was tested by Western blotting. The invasion of PANC-1 cells was measured by traswell assay. The migration of PANC-1 cells was detected by wound healing. Results Compared with control group, the level of miR-195 in miR-195 mimic group was increased （P 〈0. 001 ） . The mRNA level of VEGFA in miR-195 mimic group was lower than control group （P 〈 0. 001 ） . The luciferase assay showed that VEGFA was a direct target of has-mir- 195-5 p. The promotion of pc-VEGFA on the expression of VEGFA was repressed by miR-195 m/m-ic. The number of invasive cells per field in miR-195 mimic group was tower man control group t r 〈0.01 ） . The number of invasive cells per field in pc-VEGFA group was higher than in control group （P 〈 0. 01 ） . Compared with pc-VEGFA group, the number of invasive cells per field in miR-195 mimic ＋ pc-VEGFA group was reduced （P 〈 0. 01 ） . Compared with control group, the migration in miR-195 mimic group was decreased and migration in pe-VEGFA group was elevated （P 〈0. 01 ） . The migration in miR-195 mimic ＋ pc-VEGFA group was lower than in pc-VEGFA group （P 〈0. 01） . Conclusion has-miPt-195-5 p attenuates the in＇casion and migration of hu- man pancreatic carcinoma PANC-1 cells by targeting VEGFA.

关 键 词：胰腺癌 miR-195-5p VEGFA 侵袭 迁移 

分 类 号：R735.9[医药卫生—肿瘤]