摘要
目的初步探讨miR-146a-5p在小细胞肺癌(SCLC)耐药机制中的作用。方法通过qRT-PCR检验SCLC耐药细胞株(H69-AR)和敏感细胞株(H69)中miR-146a-5p表达水平,探究miR-146a-5p与SCLC耐药的相关性;分别通过慢病毒载体以稳定转染miR-146a-5p mimic或miR-146a-5p inhibitor使其细胞中的miR-146a-5p上调或抑制,再利用CCK-8法和划痕实验验证miR-146a-5p对SCLC细胞株的生物学影响;查阅相关文献以及应用目前生物学最权威软件预测miR-146a-5p的靶基因,应用Western blot、qRT-PCR检测miR-146a-5p与预测靶基因两者之间的关系。结果H69-AR相对H69细胞株的miR-146a-5p表达水平为(1∶22.70),差异有统计学意义(P<0.05);生物学信息数据库预测miR-146a-5p可能靶基因为白介1受体相关激酶1(IRAK1)和肿瘤坏死因子受体相关因子6(TRAF6);H69-AR相对H69细胞株中mIRAK1、mTRAF6的表达水平差异分别为(4.92∶1.00)、(2.76∶1.00),差异有统计学意义(P<0.05);H69-AR细胞株转染miR-146a-5p mimic后,与对照组比较,细胞抑制率上升,迁移速度减缓,同时mIRAK1、mTRAF6表达水平分别减少为(1.00∶14.55)、(1∶3.20),并且IRAK1、TRAF6蛋白表达也分别减少为(0.24∶1.00)、(0.31∶1.00),差异有统计学意义(P<0.05);H69细胞株转染miR-146a-5p inhibitor后,与对照组相比,细胞抑制率下降,迁移速度增快,同时mIRAK1、mTRAF6表达水平分别增加为(10.86∶1.00)、(4.64∶1.00),IRAK1、TRAF6蛋白表达也分别增加为(4.76∶1.00)、(3.96∶1.00),差异有统计学意义(P<0.05)。结论miR-146a-5p通过靶向IRAK1和TRAF6基因抑制SCLC耐药。
Objective To explore the role of miR-146 a-5 p in the drug resistance mechanism of small cell lung cancer(SCLC).Methods The expression levels of miR-146 a-5 p in drug-resistant SCLC cell lines(H69-AR)and sensitive cell lines(H69)were tested by qRT-PCR to explore the correlation between miR-146 a-5 p and SCLC drug resistance.MiR-146 a-5 p was upregulated or inhibited by lentivirus vector stably transfected with miR-146 a-5 p mimic or miR-146 a-5 p inhibitor,respectively.The biological effects of miR-146 a-5 p on SCLC cell lines were verified by CCK-8 method and Nick experiment.The target gene of mir-146 a-5 p was predicted by searching related literatures and using the most authoritative software in biology.The correlation between miR-146 a-5 p and the predicted target genes was detected by western-blot,real-time quantitative qPCR and other methods.Results The expression level of miR-146 a-5 p in H69-AR compared with H69 cell line was(1∶22.70)(P<0.05).The potential target genes of miR-146 a-5 p were predicted by biological software to be interleukin-1 receptor-related kinase 1(IRAK1)and tumor necrosis factor receptor-related factor 6(TRAF6).The expression levels of mIRAK1 and mTRAF6 in H69-AR relative to H69 cells were significantly different(4.92∶1.00 and 2.76∶1.00)(P<0.05).After transfection of miR-146 a-5 p mimic in H69 AR cell line,compared with the control group,the cell inhibition rate increased,the migration rate slowed down,and the expression levels of mIRAK1 and mTRAF6 decreased to(1.00∶14.55)and(1∶3.20),respectively.The expressions of IRAK1 and TRAF6 decreased to(0.24∶1.00)and(0.31∶1.00)(P<0.05).After transfection of miR-146 a-5 p inhibitor in H69 cell line,compared with the control group,the cell inhibition rate decreased and the migration rate increased.At the same time,the expression levels of mIRAK1 and mTRAF6 increased to(10.86∶1.00)and(4.64∶1.00),respectivelywhile the expressions of IRAK1 and TRAF6 increased to(4.76∶1.00)and(3.96∶1.00),respectively(P<0.05).Conclusion MiR-146 a
作者
梁雪
吕磊
钱立庭
李明
韩丹丹
Liang Xue;Li Lei;Qian Liting(Xinxiang Medical University,Xinxiang 453000;Anhui Prouincial Cancer Hospital Affiliated to Xinxiang Medical University,Hefei 230031;Epigenetic Laboratory of Anhui Cancer Hospital,Hefei 230031;Radiotherapy Dept,Western Region,The First Afiliated Hospital of University of Science and Technology of China,Hefei 230031)
出处
《安徽医科大学学报》
CAS
北大核心
2020年第3期333-339,共7页
Acta Universitatis Medicinalis Anhui
基金
国家自然科学基金青年项目(编号:81602230)
安徽省重点研究与开发计划项目(编号:1704a0802157)。
