微小RNA-146-5p靶向Notch2对非小细胞肺癌增殖、凋亡和侵袭的影响

Effect of microRNA-146-5p targeting Notch2 on proliferation,apoptosis and invasion of non-small cell lung cancer

目的探讨微小RNA(miRNA,miR)-146-5p靶向Notch2对非小细胞肺癌增殖、凋亡和侵袭的影响。方法选取南阳医学高等专科学校第一附属医院2018年5月至2022年5月收治的58例非小细胞肺癌和癌旁组织作为研究对象,采用荧光定量聚合酶链反应(PCR)分析肿瘤和癌旁组织miR-146-5p表达水平。采用慢病毒建立miR-146-5p过表达非小细胞肺癌A549细胞系,分别为miRNA对照组和miR-146-5p组。采用细胞计数试剂盒(CCK-8)和体外移植瘤实验分析肿瘤细胞增殖能力,采用流式细胞术分析细胞凋亡水平;采用Transwell分析细胞侵袭能力;生物信息学和双荧光素酶报告基因分析miR-146-5p靶基因。蛋白质免疫印迹分析靶基因的表达水平。组间数据比较采用t检验。结果癌旁组织中miR-146-5p表达水平(1.09±0.16)明显高于非小细胞肺癌组织表达水平(0.48±0.12),差异有统计学意义(t=23.940,P<0.05)。miRNA对照组细胞miR-146-5p表达水平(0.96±0.41)明显低于miR-146-5p组细胞(2.69±0.25),差异有统计学意义(t=8.094,P<0.05)。miRNA对照组细胞吸光度值(1.96±0.07)明显高于miR-146-5p组细胞(1.56±0.08),差异有统计学意义(t=9.123,P<0.05)。miRNA对照组细胞体内肿瘤体积[(691.50±51.29)mm3]明显高于miR-146-5p组细胞[(443.80±61.43)mm3],差异有统计学意义(t=9.787,P<0.05)。miRNA对照组细胞凋亡比例[(3.64±1.24)%]明显低于miR-146-5p组细胞[(23.26±2.91)%],差异有统计学意义(t=13.910,P<0.05)。miRNA对照组细胞侵袭数量[(98.80±14.97)%]明显低于miR-146-5p组细胞[(69.80±10.89)%],差异有统计学意义(t=3.502,P<0.05)。,Nocth2是miR-146-5p的靶基因。miRNA对照组细胞Nocth2蛋白表达水平(1.21±0.12)明显低于miR-146-5p组细胞(0.63±0.10),差异有统计学意义(t=9.282,P<0.05)。结论miR-146-5p通过调节靶蛋白Notch2表达水平,调节着非小细胞肺癌的增殖、凋亡和侵袭能力。

Objective To investigate the effects of microRNA(miRNA,miR)-146-5p targeting Notch2 on the proliferation,apoptosis and invasion of non-small cell lung cancer.Methods 58 cases of non-small cell lung cancer and adjacent tissues treated in our hospital from May 2018 to May 2022 were selected as research objects,and the expression level of miR-146-5p in tumor and adjacent tissues was analyzed by fluorescence quantitative polymerase chain reaction(PCR).Lentivirus was used to establish a non-small cell lung cancer A549 cell line with miR-146-5p overexpression,which was divided into miRNA control group and miR-146-5p group.The proliferation of tumor cells was analyzed by cell counting kit-8(CCK-8)and in vitro tumor transplantation experiments.The level of apoptosis in miRNA control group and miR-146-5p group was analyzed by flow cytometry.The invasion in miRNA control group and miR-146-5p group was analyzed by Transwell.Bioinformatics and double luciferase reporter gene analysis miR-146-5p target gene.The expression level of target gene was analyzed by Western blotting.T-test was used to compare the data between groups.Results The expression level of miR-146-5p in the adjacent tissues(1.09±0.16)was significantly higher than that in the non-small cell lung cancer tissues(0.48±0.12,t=23.940,P<0.05).The expression level of miR-146-5p in miRNA control group cells(0.96±0.41)was significantly lower than that in miR-146-5p group cells(2.69±0.25,t=8.094,P<0.05).The light absorption value of miRNA control group cells(1.96±0.07)was significantly higher than that of miR-146-5p group cells(1.56±0.08,t=9.123,P<0.05).The tumor volume in miRNA control group[(691.50±51.29)mm3]was significantly higher than that in miR-146-5p group[(443.80±61.43)mm3,t=9.787,P<0.05].The percentage of apoptosis in miRNA control group[(3.64±1.24)%]was significantly lower than that in miR-146-5p group[(23.26±2.91)%,t=13.910,P<0.05].The number of cell invasion in miRNA control group[(98.80±14.97)cells]was significantly lower than that in miR-146-5p group[(69.80±10.89)cells,t=3.502,P<0.05].Nocth2 is the target gene of miR-146-5p.The expression level of Nocth2 protein in miRNA control group cells(1.21±0.12)was significantly lower than that in miR-146-5p group cells(0.63±0.10,t=9.282,P<0.05).Conclusion MiR-146-5p plays an important role in the proliferation,apoptosis and invasion of non-small cell lung cancer by regulating the expression level of Notch2,a target protein.