血管内皮生长因子-C和人表皮因子受体2共沉默对人肺腺癌细胞生物学行为的影响

目的 观察靶向血管内皮生长因子-C(VEGF-C)、人表皮因子受体2(HER2/neu)基因的小干扰RNA(siRNA)转染对肺腺癌细胞株A549生物学行为的影响.方法 构建靶向VEGF-C、HER2/neu的siRNA表达质粒,转染A549细胞,Transwell小室检测细胞体外侵袭能力,噻唑蓝(MTY)比色法检测细胞增殖活性,流式细胞术测定凋亡率.结果 siVEGF-C组、siHER2/neu组、共转染组的细胞侵袭数分别为(42.76±2.87)、(47.10±3.63)、(40.50±3.21),显著低于阴性对照组(61.00±2.48,P＜0.01);转染48 h生长抑制率分别为(19.77±1.15)%、(22.98±2.13)%、(41.27±0.51)%,显著高于阴性对照组(4.99±0.14)%(P＜0.01);转染48 h肿瘤细胞凋亡率分别为(10.8±1.8)%、(11.7±1.5)%、(20.2±3.4)%,显著高于阴性对照组(2.6±0.7)%(P＜0.01).与单独一种siRNA转染比较,联合转染肿瘤细胞生长抑制率增加(P＜0.01),凋亡率增加(P＜0.01).结论 靶向VEGF-C、HER2/neu的siRNA转染后可以显著降低A549细胞侵袭能力,抑制增殖,诱导细胞凋亡,共转染后对增殖和凋亡影响更显著.

Effects of silence of VEGF-C and HER2/neu gene on the biologicai behaviors of human lung ade nocarcinoma cells

objective To study the effect of small interferenee RNA(siRNA)expression vector of VEGF-C and HER2/neu on the biological behaviors of human lung adenocarcinoma cell line A549.Methotis We constructed the eukaryotie expression vector of siRNA targeting VEGF-C and HER2/neu gene,and transfected A549 cells respectively.The invasion ability of the tumor cells was detected by Transwell cell culture chamber,the proliferation activity was determined by MTT assay,and the apoptosis rate was determined by flow cytometry.Results In siVEGF-C,siHER2/neu,and combined groups,the number of invasion cells was(42.76±2.87),(47.10±3.63),and(40.50±3.21)respectively,which was reduced obviously as compared with(61.00±2.48)in the negative control group(P＜0.01).The inhibifive ratio of eell proliferation at48 h was(19.77±1.15)%,(22.98±2.13)%,(41.27±0.51)%in siVEGF-C,siHER2/neu,and combined groups respectively,which was increased significantly as compared with(4.99±0.14)%in the negative control group(P＜0.01).The apoptosis rate at48 h was(10.8±1.8)%,(11.7±1.5)%,(20.2±3.4)%siVEGF-C,siHER2/neu,and combined groups respectively,which wilts increased as compared with(2.6±0.7)%in the negative control group(P＜0.01).As compared with siVEGF-C and siHER2/neu groups.the inhibition rate of cells proliferation and apeptosis rate were increased in combined group(P＜0.01).Conclusion The siRNA vectors targeting VEGF-C and HER2/neu genes could efficiently inhibit the invasion and proliferation ability,and induce the apoptosis of A549 cells as well.Besides,co-transfection of VEGF-C and HER2/neu gene has a more markedly effect on the proliferation and apoptosis.