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The increased application of graphene oxide (GO), a new carbon-based engineered nanomaterial, has generated a potential toxicity in humans and the environment. Previous studies have identified some dysregulated microRNAs (miRNAs), such as up-regulated  mir-235 , in organisms exposed to GO. However, the detailed mechanisms of the dysregulation of miRNA underlying GO toxicity are still largely elusive. In this study, we employed  Caenorhabditis elegans  as an in vivo model to investigate the biological function and molecular basis of  mir-235  in the regulation of GO toxicity. After low concentration GO exposure,  mir-235  ( n4504 ) mutant nematodes were sensitive to GO toxicity, implying that  mir-235  mediates a protection mechanism against GO toxicity. Tissue-specific assays suggested that  mir-235  expressed in intestine is required for suppressing the GO toxicity in  C. elegans .  daf-12 , a gene encoding a member of the steroid hormone receptor superfamily, acts as a target gene of  mir-235  in the nematode intestine in response to GO treatment, and RNAi knockdown of  daf-12  suppressed the sensitivity of  mir-235 ( n4503 ) to GO toxicity. Further genetic analysis showed that DAF-12 acted in the upstream of DAF-16 in insulin/IGF-1 signaling pathway and PMK-1 in p38 MAPK signaling pathway in parallel to regulate GO toxicity. Altogether, our results revealed that  mir-235  may activate a protective mechanism against GO toxicity by suppressing the DAF-12-DAF-16 and DAF-12-PMK-1 signaling cascade in nematodes, which provides an important molecular basis for the in vivo toxicity of GO at the miRNA level.

The C. elegans miR-235 regulates the toxicity of graphene oxide via targeting the nuclear hormone receptor DAF-12 in the intestine