摘要
目的:研究沉默长链非编码RNA DGCR5对肺腺癌细胞增殖、侵袭、迁移及Wnt/β-catenin信号通路的影响,探讨DGCR5对肺腺癌的可能作用机制。方法:将H1650和A549细胞株分为空白对照组(Blank)、阴性对照组(NC)和siRNA-DGCR5组(si-DGCR5),si-DGCR5组细胞转染siRNA-DGCR5,NC组细胞转染siRNA-NC,Blank组不转染。qRT-PCR测量细胞中DGCR5水平,CCK8检测细胞增殖(OD值),Transwell测定细胞侵袭能力,划痕实验测定细胞迁移能力,Western blot测定各组H1650和A549细胞中E-cadherin、Vimentin、β-catenin、磷酸化糖原合成酶激酶-3(p-gsk-3β)蛋白水平。结果:H1650和A549细胞中,与Blank组和NC组相比,si-DGCR5组DGCR5水平、细胞OD值、穿膜细胞数、细胞迁移距离降低(P<0.05),E-cadherin、p-gsk-3β蛋白水平升高(P<0.05),Vimentin、β-catenin蛋白水平降低(P<0.05);Blank组和NC组DGCR5水平、细胞OD值、穿膜细胞数、细胞迁移距离及细胞E-cadherin、Vimentin、β-catenin、p-gsk-3β蛋白水平差异无统计学意义(P>0.05)。结论:沉默DGCR5可能通过抑制Wnt/β-catenin信号通路抑制上皮间质转化,从而抑制肺腺癌细胞增殖、侵袭和迁移。
Objective:To investigate the effects of silencing long non-coding RNA DGCR5 on proliferation,invasion,migration and Wnt/β-catenin signaling pathway in lung adenocarcinoma cells,and to explore possible mechanism of DGCR5 on lung adenocarcinoma.Methods:H1650 and A549 cell beads were divided into blank control group(Blank),negative control group(NC)and siRNA-DGCR5 group(si-DGCR5).Cells of si-DGCR5 group were transfected with siRNA-DGCR5,cells of NC group were transfected with siRNA-NC,cells of Blank group were not transfected.qRT-PCR was used to measure DGCR5 levels in cells.CCK8 was used to measure proliferation ability(OD)of cells,Transwell was used to measure cell invasiveness,scratch test was used to determine cell migration ability.Western blot was used to determine the E-cadherin,Vimentin,β-catenin and phosphorylated glycogen synthase kinase-3(p-gsk-3β)protein levels in H1650 and A549 cells.Results:In H1650 and A549 cells,compared with Blank and NC group,the DGCR5 level,cell OD value,transmembrane cell number and cell migration distance were decreased(P<0.05),E-cadherin and p-gsk-3βprotein levels were increased(P<0.05),Vimentin andβ-catenin protein levels were decreased(P<0.05)in the si-DGCR5 group.There were no significant differences in DGCR5 levels,cell OD values,transmembrane cell numbers,cell migration distance and the protein levels of E-cadherin,Vimentin,β-catenin,and p-gsk-3βbetween Blank group and NC group(P>0.05).Conclusion:Silencing DGCR5 may inhibit epithelial-mesenchymal transition by inhibiting Wnt/β-catenin signaling pathway,thereby inhibiting proliferation,invasion and migration of lung adenocarcinoma cells.
作者
郭水根
舒慧珍
GUO Shui-Gen;SHU Hui-Zhen(Department of Respiratory,Affiliated Pudong Hospital of Fudan University,Shanghai 201399,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2020年第23期2866-2871,共6页
Chinese Journal of Immunology
