miR-125a靶向转录无调性碱性螺旋环螺旋转录因子8对肺腺癌恶性进展的影响

miR-125a targeting transcription factor atonal homolog 8 is involved in the progression of lung adenocarcinoma

目的探讨miR-125a通过靶向转录无调性碱性螺旋环螺旋转录因子8(ATOH8)对肺腺癌恶性进展的影响。方法通过在线数据库UALCAN分析ATOH8在肺腺癌中的表达水平及其与患者生存率的相关性,以及miR-125a在肺腺癌中的表达水平及其与肺癌进展的关系;提取肺腺癌及癌旁正常组织总RNA,实时PCR分析ATOH8表达水平差异;将ATOH8过表达载体转染至肺腺癌细胞中,CCK-8法检测过表达ATOH8对肺腺癌细胞存活能力的影响;预测与ATOH8的3’非翻译区(UTR)结合的微RNA(miRNA),将miR-125a模拟物和抑制物转染至肺腺癌细胞中,实时PCR和Western blotting检测ATOH8的表达水平变化。结果数据库及实时PCR验证结果显示,ATOH8在肺腺癌组织中显著下调(P<0.01);过表达ATOH8能够显著降低肺腺癌细胞存活能力(P<0.05);高表达ATOH8组患者5年生存率显著升高(P<0.05);miR-125a能够与ATOH8的3’UTR结合,显著抑制其表达(P<0.05)。miR-125a在有吸烟史、中晚期和淋巴转移的肺腺癌患者中显著上调(P<0.05)。结论ATOH8是肺腺癌潜在的抑癌基因,能够抑制肺腺癌细胞存活的能力,影响患者的5年生存率。miR-125a表达水平与吸烟史、肿瘤分期和淋巴转移密切相关。miR-125a能够靶向抑制肺腺癌患者ATOH8表达,是ATOH8在肺腺癌中异常表达的潜在因素。

Objective To investigate the effect of transcription factor atonal homolog 8(ATOH8)and miR-125a on lung cancer progression and its potential upstream regulatory mechanism.Methods ATOH8 expression levels in lung adenocarcinoma and their correlation with survival rate were analyzed using the online database UALCAN.miR-125a expression levels in lung adenocarcinoma and their relationship with lung cancer progression were also analyzed using the UALCAN database.Total RNA extracted from lung adenocarcinoma tumors and adjacent normal tissues was used to perform real-time PCR in order to analyze these expression levels.The effect of ATOH8 overexpression on lung adenocarcinoma cell survival was detected using CCK-8 assays.A miR-125a mimic and inhibitor were transfected into lung adenocarcinoma cells,and ATOH8 expression levels were detected by real-time PCR and Western blotting.Results Statistical analysis showed that ATOH8 was significantly down-regulated in lung adenocarcinoma tissues(P<0.01)and ATOH8 overexpression significantly reduced the survival of lung adenocarcinoma cells(P<0.05).Furthermore,the five-year survival rate of patients with high ATOH8 expression levels was significantly increased(P<0.05).miR-125a can bind to the 3’untranslated regions(3’UTR)of ATOH8 and significantly inhibit its expression levels(P<0.05).However,miR-125a was significantly up-regulated in lung adenocarcinoma patients with a history of smoking,middle and advanced stage,and lymphatic metastasis(P<0.05).Conclusion ATOH8,as a potential tumor suppressor gene,can inhibit lung adenocarcinoma cell survival and affect the five-year survival rate of patients.miR-125a expression levels were closely related to smoking history,tumor stage,and lymphatic metastasis.Overall,the inhibiting effect of miR-125a against ATOH8 is a potential reason for abnormal ATOH8 expression in lung adenocarcinoma.