circBMPR2竞争性结合miR-6814-5p抑制肺腺癌细胞的作用机制  

circBMPR2 inhibits the proliferation,migration,and invasion of lung adenocarcinoma cells by competitively binding to miR-6814-5p

作　　者：王慧敏 邹晓莉 曹碧月 王雪君 王亚东 严玉兰[1] WANG Hui-min;ZOU Xiao-li;CAO Bi-yue;WANG Xue-jun;WANG Ya-dong;YAN Yu-lan(Department of Respiratory,the Affiliated People's Hospital of Jiangsu University,Zhenjiang 212002,Jiangsu,China;Department of Clinical Medicine,Jiangsu University School of Medicine,Zhenjiang 212013,Jiangsu,China)

机构地区：[1]江苏大学附属人民医院呼吸科,镇江212002 [2]江苏大学医学院,镇江212013

出　　处：《医学研究生学报》2022年第7期694-700,共7页Journal of Medical Postgraduates

基　　金：江苏省自然科学基金(BK20151333);镇江市重点研发计划项目(SH2021032)。

摘　　要：目的环状RNA(circRNA)参与肺腺癌的发生发展过程,然而作用机制有待进一步研究。文章旨在探究circBMPR2对肺腺癌细胞增殖、迁移、侵袭能力的影响及调控机制。方法收集30对肺腺癌组织和癌旁组织,其中5对标本进行高通量测序,根据高通量测序结果选取显著差异性表达的circBMPR2。利用qRT-PCR检测25对肺腺癌组织及癌旁组织、肺腺癌细胞系(A549、H1975、PC9)和正常支气管黏膜上皮细胞(BEAS-2B)中circBMPR2的表达水平。过表达质粒转染PC9细胞;小干扰RNA转染A549细胞,共分为两组对照,分别是PC9细胞OE-circ及对照组circ-NC,A549细胞si-circ及对照组si-NC。利用平板克隆形成实验、细胞划痕实验、Transwell实验检测细胞的增殖、迁移和侵袭能力。用蛋白质印迹法检测上皮间质转化(EMT)相关蛋白(E-cadherin、N-cadherin、Vimentin)的表达水平。通过双荧光素酶实验及qRT-PCR验证circBMPR2与miR-6814-5p的靶向关系。通过蛋白质印迹法验证ARID1A是miR-6814-5p靶基因。结果circBMPR2在肺腺癌组织(0.30±0.10)中的表达水平较癌旁组织(1.00±0.29)低(P<0.05)。与正常支气管黏膜上皮细胞相比,3株肺腺癌细胞株中的circBMPR2相对表达水平均低(P<0.05)。与circ-NC组相比,OE-circ组PC9细胞克隆形成率、细胞迁移率、侵袭细胞数下降且N-cadherin、Vimentin蛋白相对表达量低(P<0.05),E-cadherin蛋白相对表达量高(P<0.05)。与si-NC组相比,si-circ组A549细胞细胞克隆形成率、细胞迁移率、侵袭细胞数上升且N-cadherin、Vimentin蛋白相对表达量高(P<0.05),E-cadherin蛋白相对表达量低(P<0.05)。qRT-PCR实验表明,PC9细胞中OE-circ组miR-6814-5p相对表达水平(0.47±0.04)较circ-NC组(1.00±0.16)降低(P<0.05);A549细胞中si-circ组miR-6814-5p相对表达水平(2.51±0.60)较si-NC组(1.00±0.12)明显升高(P<0.05)。结论circBMPR2通过竞争性结合miR-6814-5p促进ARID1A的表达,抑制肺腺癌细胞的增殖、迁移和侵袭能�Objective Existing studies show that circular RNA(circRNA)participates in the development of lung adenocarcinoma,but the underlying mechanism needs further study.This study aimed to investigate the effect of circBMPR2 on the proliferation,migration and invasion capacity of lung adenocarcinoma cells and its regulatory mechanism.Methods 30 pairs of lung adenocarcarcinoma and parparcancer tissues were collected,5 samples were used for high-throughput sequencing and the differential expression circBMPR2 was selected according to the results.The expression of circBMPR2 in 25 lung adenocarcarcinoma tissues and corresponding normal adjacent lung tissues,lung adenocarcinoma cell lines(A549,H1975,PC9)and normal bronchial mucosal epithelial cells(BEAS-2B)were detected by qRT-PCR.Overexpressed plasmids were transfected into PC9 cells and siRNA were transfected into A549 cells,two control groups,PC9 cell OE-circ group and control group circ-NC,A549 cell si-circ and control group si-NC were divided.The proliferation,migration and invasion ability of cells were detected by plate cloning formation assay,scratch test and Transwell assay,respectively.Expression levels of epithelial mesenchymal transition(EMT)associated protein(E-cadherin,N-cadherin,Vimentin)were detected by Western blot.The targeting relationship between circBMPR2 and miR-6814-5p was verified by dual luciferase experiments and qRT-PCR.ARID1A was verified as a miR-6814-5p target gene by Western blot.Results The circBMPR2 expression level in lung adenocarcinoma tissues(0.30±0.10)was lower than that in adjacent lung tissues(1.00±0.29).The circBMPR2 expression levels in three lung adenocarcinoma cell lines were lower than that in normal bronchial mucosal epithelial cells.Compared with the circ-NC group,clone formation rate,cell mobility,invasion cell number,N-cadherin,Vimentin protein relative expression of OE-circ group PC9 cell were significantly decreased,while the relative expression of E-cadherin protein was increased.Compared with the si-NC group,clone format

关 键 词：肺腺癌 circBMPR2 miR-6814-5p ARID1A 细胞生物学行为 

分 类 号：R734.2[医药卫生—肿瘤]