Lei Zhentao, Lv Yuan, Wang Weijie, Guo Qin, Zou Feifei, Hu Shengli, Fang Fujin, Tian Mengmeng, Liu Bingqian, Liu Xianmiao, Ma Kai, Ma Lei, Zhou Dan, Zhang Donghui, Sun Yan, Shen Bo, Zhu Changliang
Department of Pathogen Biology, Nanjing Medical University, Nanjing, China.
Parasitol Res. 2015 Feb;114(2):699-706. doi: 10.1007/s00436-014-4236-7. Epub 2014 Nov 26.
MicroRNAs (miRNAs) regulate gene expression and biological processes including embryonic development, innate immunity, and infection in many species. Emerging evidence indicates that miRNAs are involved in drug resistance. However, little is known about the relationship between the miRNAs and insecticide resistance in mosquitos. Here, we reported that conserved miR-278-3p and its target gene are critical for pyrethroid resistance in Culex pipiens pallens. We found that CYP6AG11 is the target of miR-278-3p, through bioinformatic analysis and experimental verification. The expression level of miR-278-3p was lower, whereas the level of CYP6AG11 was higher in deltamethrin-resistant strain, which were detected using quantitative reverse transcription PCR (qRT-PCR). We also found that CYP6AG11 was regulated by miR-278-3p via a specific target site with the 3' untranslated region (UTR) by luciferase reporter assay. In addition, overexpression of CYP6AG11 in the mosquito C6/36 cells showed better proliferation than the cells with empty vector when treated by deltamethrin at different concentrations. Moreover, the overexpression of miR-278-3p through microinjection led to a significant reduction in the survival rate, and the level of CYP6AG11 was simultaneously reduced. These results indicated that miR-278-3p could regulate the pyrethroid resistance through CYP6AG11.
微小RNA(miRNA)可调节基因表达和生物过程,包括许多物种的胚胎发育、天然免疫和感染。新出现的证据表明,miRNA与耐药性有关。然而,关于miRNA与蚊子抗杀虫剂性之间的关系却知之甚少。在此,我们报告保守的miR-278-3p及其靶基因对淡色库蚊的拟除虫菊酯抗性至关重要。通过生物信息学分析和实验验证,我们发现CYP6AG11是miR-278-3p的靶标。使用定量逆转录PCR(qRT-PCR)检测发现,在溴氰菊酯抗性品系中,miR-278-3p的表达水平较低,而CYP6AG11的水平较高。我们还通过荧光素酶报告基因测定发现,miR-278-3p通过与3'非翻译区(UTR)的特定靶位点调控CYP6AG11。此外,在不同浓度溴氰菊酯处理下,在蚊子C6/36细胞中过表达CYP6AG11比空载体细胞表现出更好的增殖能力。而且,通过显微注射过表达miR-278-3p导致存活率显著降低,同时CYP6AG11的水平也降低。这些结果表明,miR-278-3p可通过CYP6AG11调节拟除虫菊酯抗性。