RNA干扰VEGFR-2表达对Calu-1细胞增殖、迁移、侵袭力及放射效应影响  

Effects of RNA interference-mediated silencing of vascular endothelial growth factor receptor-2 on proliferation, migration, invasion, and radiation-induced effects in Calu-1 cells

作　　者：刘毅[1] 刘亮[1] 胡晨曦[2] 周莉华[2] 乔云[2] 王磊[2] 刘彬[1] 陈晖[1] 蒋晓东[2] 

机构地区：[1]徐州医学院研究生院,221000 [2]连云港市第一人民医院放疗科,222002

出　　处：《中华放射肿瘤学杂志》2015年第6期714-718,共5页Chinese Journal of Radiation Oncology

基　　金：国家自然科学基金（81472792）;卫生部课题基金（W201210）;江苏省自然科学基金（BK2012661）

摘　　要：目的：研究VEGFR？2对肺癌细胞系Calu？1细胞增殖、迁移、侵袭及联合放射后凋亡率影响，并探讨其可能机制。方法 siRNA敲低Calu？1细胞中的VEGFR？2基因，借助实时荧光定量PCR和蛋白印迹法检测VEGFR？2表达水平的变化；将细胞分为对照组、VEGF组、VEGFR？2基因敲低组和基因敲低加VEGF组。利用CCK8法、细胞划痕实验及Transwell实验分别检测细胞增殖、迁移和侵袭能力变化，利用蛋白印迹法检测VEGFR？2及下游相关信号通路蛋白表达水平变化；各组细胞联合放射后，检测细胞凋亡。结果 RNA干扰VEGFR？2后Calu？1细胞中VEGFR？2的mRNA水平和蛋白水平均降低（ P＝0．001、0．000）；RNA干扰VEGFR？2后Calu？1细胞的增殖、迁移、侵袭能力均降低（ P＝0．000、0．000、0．031）；RNA干扰VEGFR？2后Calu？1细胞中Akt、ERK 1／2、p38的蛋白磷酸化水平均降低（ P＝0．336、0．986、0．553）；RNA干扰VEGFR？2后联合放射后Calu？1细胞的凋亡率增加（ P＝0．012），RNA干扰VEGFR？2后HIF？1α蛋白表达被抑制（ P＝0．016）。结论 VEGFR？2基因表达敲低后显著抑制了Calu？1细胞的多项生理功能，并提高了放射后细胞凋亡率。Objective To investigate the effects of vascular endothelial growth factor receptor？2 （ VEGFR？2） on proliferation, migration, invasion, and apoptosis after radiotherapy in lung cancer cell line Calu？1, and to explore the probable mechanisms. Methods Small interference RNA （ siRNA ）？mediated silencing of VEGFR？2 gene was performed on Calu？1 cells, and the mRNA and protein expression of VEGFR？2 was determined by quantitative real？time PCR and Western blot, respectively. The cells were divided into control group, vascular endothelial growth factor （ VEGF ） group, VEGFR？2 specific siRNA （siKDR） group, and siKDR＋VEGF group. The changes in proliferation, migration, and invasion were evaluated by the CCK8 assay, cell scratch wound？healing assay, and transwell migration assay, respectively. The protein expression of VEGFR？2 and proteins in the related downstream signaling pathway was measured by Western blot. Apoptosis in each group was determined after radiotherapy. Results After RNA interference？mediated silencing of VEGFR？2, the mRNA and protein expression of VEGFR？2 was significantly reduced （ P=0. 001,P=0. 000）;the proliferation, migration, and invasion of Calu？1 cells were also significantly reduced （ P=0. 000,P=0. 000,P=0. 000）;the phosphorylation levels of AKT, ERK 1/2, and p38 were significantly reduced in Calu？1 cells （ P=0. 336,P=0. 986,P=0. 553）;the apoptosis in Calu？1 cells was significantly elevated （ P=0. 0012）;the protein expression of HIF？1α was significantly inhibited （ P= 0. 016 ） . Conclusions The VEGFR？2 gene silencing significantly inhibits several physiological functions of Calu？1 cells and elevates the apoptosis rate after radiotherapy.

关 键 词：小分子干扰核糖核酸 血管内皮生长因子受体2 Calu-1细胞系 放射效应 

分 类 号：R734.2[医药卫生—肿瘤]