Enterovirus 71 (
EV
71) is the major causative agent of hand‐foot‐and‐mouth disease in young children and can cause severe cerebral and pulmonary complications and even fatality. This study aimed at elucidating whether and how
EV
71 infection is regulated by a cellular micro
RNA
, miR‐127‐5p. We found that miR‐127‐5p can downregulate the expression of
SCARB
2, a main receptor of
EV
71, by targeting two potential sites in its 3′
UTR
region and inhibit
EV
71 infection. Meanwhile, miR‐127‐5p expression was upregulated during
EV
71 infection. Notably, transfecting cells with miR‐127‐5p mimics led to a significant decrease in viral replication, while inhibition of endogenous miR‐127‐5p facilitated viral replication. Furthermore, our evidence showed that miR‐127‐5p did not affect postentry viral replication. Taken together, these results indicated that miR‐127‐5p inhibited
EV
71 replication by targeting the
SCARB
2
mRNA
.
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