摘要
目的 探讨LncRNA LINC01857调节miR-199a-3p/肝源性生长因子(HDGF)轴对肺腺癌A549细胞顺铂(DDP)化学敏感性的影响。方法 将体外培养的人肺腺癌耐DDP细胞株(A549/DDP)随机分为NC组(正常培养的A549/DDP细胞)、si-NC组、si-LINC01857组、miR-NC组、miR-199a-3p mimic组、si-LINC01857+inhibitor NC组、si-LINC01857+miR-199a-3p inhibitor组。qRT-PCR检测LINC01857、miR-199a-3p、HDGF在肺腺癌组织、不同细胞及各组A549/DDP细胞中的表达水平;检测A549/DDP细胞增殖、迁移、侵袭及凋亡情况;Western Blot检测Bax、Bcl-2表达;验证miR-199a-3p与LINC01857、HDGF的关系。结果 肺腺癌组织中LINC01857、HDGF mRNA表达高于癌旁组织,miR-199a-3p低于癌旁组织(P<0.05)。BEAS-2B细胞、A549细胞、A549/DDP细胞中LINC01857、HDGF mRNA表达均依次升高,miR-199a-3p依次降低(P<0.05)。si-LINC01857组LINC01857、HDGF mRNA表达、A549/DDP细胞增殖率、划痕愈合率、侵袭细胞数及Bcl-2表达均低于NC组和si-NC组,miR-199a-3p表达、A549/DDP细胞凋亡率及Bax表达均高于NC组和si-NC组(P<0.05);miR-199a-3p mimic组LINC01857、HDGF mRNA表达、A549/DDP细胞增殖率、划痕愈合率、侵袭细胞数及Bcl-2表达均低于miR-NC组,miR-199a-3p表达、A549/DDP细胞凋亡率及Bax表达均高于miR-NC组(P<0.05);si-LINC01857+miR-199a-3p inhibitor组LINC01857、HDGF mRNA表达、A549/DDP细胞增殖率、划痕愈合率、侵袭细胞数及Bcl-2表达均高于si-LINC01857+inhibitor NC组,miR-199a-3p表达、A549/DDP细胞凋亡率及Bax表达均低于si-LINC01857+inhibitor NC组(P<0.05)。双荧光素酶报告基因实验、RNA免疫沉淀、RNA pull-down实验验证miR-199a-3p和LINC01857、HDGF均存在靶向调控关系(P<0.05)。结论 下调LINC01857表达可抑制A549/DDP细胞增殖、迁移和侵袭,诱导其凋亡,增强其DDP化学敏感性,可能与调控miR-199a-3p/HDGF轴有关。
Objective To investigate the effects of LncRNA LINC01857 on cisplatin(DDP)chemotherapy sensitivity of lung adenocarcinoma A549 cells by regulating the miR-199a-3p/hepatoma derived growth factor(HDGF)axis.Methods Human lung cancer DDP resistant cell lines(A549/DDP)cultured in vitro were randomly separated into NC group(normal cultured A549/DDP cells),si-NC group,si-LINC01857 group,miR-NC group,miR-199a-3p mimic group,si-LINC01857+inhibitor NC group,and si-LINC01857+miR-199a-3p inhibitor group.qRT-PCR was applied to detect the expression levels of LINC01857,miR-199a-3p,and HDGF in lung cancer tissue,different cells,and A549/DDP cells in each group.The proliferation,migration,invasion,and apoptosis of A549/DDP cells were detected.Western blot was applied to detect the expression of Bax and Bcl-2.The relationship between miR-199a-3p and LINC01857 and HDGF was validated.Results The expression of LINC01857 and HDGF mRNA in lung cancer tissue was higher than that in adjacent tissues,and miR-199a-3p was lower than that in adjacent tissues(P<0.05).The mRNA expression of LINC01857 and HDGF increased sequentially in BEAS-2B cells,A549 cells,and A549/DDP cells,miR-199a-3p decreased sequentially(P<0.05).The expression of LINC01857,HDGF mRNA,A549/DDP cell proliferation rate,scratch healing rate,number of invasive cells,and expression of Bcl-2 in the si-LINC01857 group were lower than those in the NC and si-NC groups,the expression of miR-199a-3p,apoptosis rate of A549/DDP cells,and the expression of Bax were higher(P<0.05).The expression of LINC01857,HDGF mRNA,A549/DDP cell proliferation rate,scratch healing rate,number of invasive cells,and expression of Bcl-2 in the miR-199a-3p mimic group were lower than those in the miR-NC group,the expression of miR-199a-3p,apoptosis rate of A549/DDP cells,and the expression of Bax were higher(P<0.05).The expression of LINC01857,HDGF mRNA,A549/DDP cell proliferation rate,scratch healing rate,number of invasive cells,and expression of Bcl-2 in the si-LINC01857+miR-199a-3p inhibitor group were higher than those in the si-LINC01857+inhibitor NC group,the expression of miR-199a-3p,apoptosis rate of A549/DDP cells,and expression of Bax were all lower(P<0.05).Dual luciferase reporter gene experiment,RNA immunoprecipitation and RNA pull-down assay confirmed the targeted regulatory relationship between miR-199a-3p with LINC01857 and HDGF(P<0.05).Conclusion Downregulation of LINC01857 expression can inhibit the proliferation,migration,and invasion of A549/DDP cells,induce their apoptosis,enhance their DDP chemotherapy sensitivity,and may be related to the regulation of the miR-199a-3p/HDGF axis.
作者
骆晴
刘明伟
LUO Qing;LIU Mingwei(Department of Thoracic Surgery,Baoji Central Hospital,Baoji,Shanxi 721008,China)
出处
《临床肺科杂志》
2025年第6期817-825,共9页
Journal of Clinical Pulmonary Medicine
