hsa_circ_0026782对肺腺癌细胞的增殖、迁移及凋亡作用  

Effects of hsa_circ_0026782 on proliferation,migration and apoptosis of lung adenocarcinoma cells

作　　者：邹晓莉 王慧敏 曹碧月 谷文露 袁海涛 严玉兰[2] ZOU Xiao-li;WANG Hui-min;CAO Bi-yue;GU Wen-lu;YUAN Hai-tao;YAN Yu-lan(School of Medicine,Jiangsu University,Zhenjiang 212013,Jiangsu,China;Department of Respiratory Medicine,Affiliated Peopled Hospital of Jiangsu University,Zhenjiang 212002,Jiangsu,China)

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

出　　处：《医学研究生学报》2021年第12期1244-1251,共8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(SH2018059)。

摘　　要：目的环状RNA(circRNA)可调节肿瘤细胞的增殖、侵袭和转移,而影响肿瘤患者预后,而环状RNA(circRNA)在肺腺癌中作用的机制尚不清楚。文章探讨环状RNA hsa_circ_0026782在肺腺癌患者中的差异表达及对肺腺癌细胞的增殖、迁移及凋亡的影响。方法通过高通量测序检测肺腺癌组织及癌旁组织中有显著差异的环状RNA hsa_circ_0026782;通过实时定量聚合酶链式反应(qRT-PCR)技术检测肺腺癌组织与癌旁组织、肺癌患者血清、术后血清与健康体检人、肺腺癌细胞系(PC9,A549,H1975)及正常细胞系(BEAS-2B)之间hsa_circ_0026782的表达水平。选择下调最为显著的肺腺癌A549细胞进行过表达质粒转染(circ_0026782),选择下调相对较少的PC9细胞进行siRNA敲减(si-circ_0026782),使用实时定量聚合酶链式反应(qRT-PCR)技术检测过表达组(circ_0026782)与空质粒组(Empty-vector)、敲减组(si-circ_0026782)与对照组(si-NC)效率;通过对肺腺癌细胞进行平板细胞克隆形成实验、CCK-8(Cell-Counting Kit-8)实验检测hsa_circ_0026782对肺腺癌细胞的增殖作用的影响;通过划痕实验和Transwell迁移实验检测hsa_circ_0026782对肺腺癌细胞迁移的作用;通过qRT-PCR检测CMTM4 mRNA表达水平,通过蛋白免疫印迹实验(Western-Blot)检测增殖相关标志物、凋亡相关标志物及目的蛋白CMTM4表达水平。结果相对于空载体组(1.19±0.52),转染过表达质粒组(534.2±39.12)能明显提高hsa_circ_002678的表达;相对于对照组(si-NC)(1.06±0.13),用小干扰RNA敲减hsa_circ_0026782(si-circ_0026782)(0.052±0.004)能明显减少hsa_circ_0026782的表达。平板克隆实验、CCK-8实验及Western blot实验结果均表明,hsa_circ_0026782的表达上调可明显抑制肺腺癌细胞增殖,敲减hsa_circ_0026782的表达下调可明显促进肺腺癌细胞的增殖D过表达hsa_drc_0026782(circ_0026782)可促进肺腺癌细胞凋亡,敲减hsa_drc_0026782(si-circ_0026782)可抑制肺腺癌细胞凋亡。细胞�Objective Lung adenocarcinoma is the most deadly malignancy worldwide.Circular RNA(circRNA)regulates the proliferation,invasion and metastasis of tumor cells,thus affects the prognosis of tumor patients.And the mechanism of the role of circular RNA(circRNA)in lung adenocarcinoma is not clear.Objective To investigate the differential expression of circrRNA hsa_circ_0026782 in patients with lung adenocarcinoma and its effect on the proliferation,migration and apoptosis of lung adenocarcinoma cells.Methods The circRNA hsa_circ_0026782 in lung adenocarcinoma and paracancerous tissues were detected bv high-throughput sequencing.Real-time quantitative polymerase chain reaction(qRT-PCR)technique was used to detect the expression levels of hsa_circ_0026782 in lung adenocarcinoma tissue and paracancerous tissue,serum of lung cancer patients,postoperative serum and healthy persons,lung adenocarcinoma cell lines(PC9,A549,H1975)and normal cell lines(BEAS-2b).Lung adenocarcinoma A549 cells with the most significant down-regulation were selected for overexpression plasmid transfection(circ_0026782),and PC9 cells with relatively less down-regulation were selected for siRNA knockdown(si-circ_0026782).The efficiency of the overexpression group(circ_0026782)versus the empty plasmid group(Empty-vector)and the knockdown group(si-circ_0026782)versus the control group(si-NC)were measured using quantitative real-time PCR(qRT-PCR).The proliferation effect of hsa_circ_0026782 on lung adenocarcinoma cells was detected by plate cell cloning and CCK-8(cell-counting kit-8)experiment.The effect of hsa_circ_0026782 on the migration of lung adenocarcinoma cells was detected by scratch assay and Transwell migration assay.CMTM4 mRNA expression was detected by qRT-PCR.The proliferation-related markers,apoptosis-related markers and CMTM4 protein expression levels were detected by Western-blot assay.Results The expression of hsa_circ_0026782 was decreased in lung adenocarcinoma tissues compared with paracancerous tissues,lung adenocarcinoma serum co

关 键 词：肺腺癌 hsa_circ_0026782 细胞增殖 细胞迁移 

分 类 号：R734.2[医药卫生—肿瘤]