Abstract

Micro RNAs (miRNAs) are major players in cellular responses to xenobiotic compounds and toxins. However, their functions in organophosphate-induced cytotoxicity remain unclear. This study investigated the involvement of miR-96-5p in the non-cholinergic toxicity of malathion in normal human kidney cells (HK-2 cells). Malathion decreased HK-2 cell viability and the expression of miR-96-5p in a dose- and time-dependent manner. In addition, transfection with miR-96-5p mimics attenuated malathion-induced HK-2 cell apoptosis, whereas transfection with a miR-96-5p inhibitor increased HK-2 cell apoptosis. Luciferase assays indicated that miR-96-5p could bind directly to the 3′-untranslated region of DDIT3, a well-known marker of endoplasmic reticulum stress. Further analyses of the expression of apoptosis-related genes and proteins indicated that miR-96-5p may function to reduce malathion-induced HK-2 cell apoptosis via regulation of the DDIT3/B-cell lymphoma (BCL)-2/caspase-3 signaling pathway. In summary, the results of the present study indicate that miR-96-5p protects HK-2 cells from malathion-induced ER stress-dependent apoptosis by targeting DDIT3.

摘要

微小RNA（miRNA）是细胞对异种化合物和毒素的反应的主要参与者。然而，它们在有机磷酸酯诱导的细胞毒性中的功能仍不清楚。这项研究调查了miR-96-5p在正常人肾细胞（HK-2细胞）中马拉硫磷的非胆碱能毒性中的作用。马拉硫磷以剂量和时间依赖性方式降低HK-2细胞的活力和miR-96-5p的表达。另外，用miR-96-5p转染可减弱马拉硫磷诱导的HK-2细胞凋亡，而用miR-96-5p抑制剂转染可增加HK-2细胞凋亡。萤光素酶分析表明，miR-96-5p可以直接结合DDIT3的3'-非翻译区，DDIT3是内质网应激的著名标记。对凋亡相关基因和蛋白表达的进一步分析表明，miR-96-5p可能通过调节DDIT3 / B细胞淋巴瘤（BCL）-2 / caspase-3来减少马拉硫磷诱导的HK-2细胞凋亡。信号通路。总之，本研究的结果表明，miR-96-5p通过靶向DDIT3保护HK-2细胞免受马拉硫磷诱导的ER应激依赖性细胞凋亡。