Acute kidney injury (AKI) is a common kidney disorder that affects public health and the incidence of AKI. Sepsis, acute ischemia or hypoxia is the main reason for the occurrence of AKI. Recently, noncoding RNA that include microRNA and long noncoding RNA (lncRNAs) were reported to play important roles in AKI as well as have the potential to serve as a biomarker or therapeutic target for the development of the diagnostic and prognostic strategies of AKI. In the current study, we aimed to investigate the expression and biological function of lncRNA nuclear enriched abundant transcript 1 (NEAT1) in ischemia‐induced AKI in patients' sample and in vitro. The expressions of NEAT1 and miR‐27a‐3p in ischemia/reperfusion‐induced AKI patients were examined by quantitative reverse transcription polymerase chain reaction. Cell injury was induced by treatment of human kidney tubular cells (HK‐2) with CoCl₂. After treatment, the influences of NEAT1 and miR‐27a‐3p on the cell apoptosis in the CoCl₂‐stimulated HK‐2 were tested by flow cytometry. The flow analysis results showed that the expression of NEAT1 was markedly higher in the ischemia‐induced AKI patients compared with normal control. Moreover, repression the expression of NEAT1 decreased CoCl₂‐induced injury in HK‐2. The expression of miR‐27a‐3p was negatively regulated by NEAT1. Inhibition the expression of NEAT1 attenuated overexpression of miR‐27a‐3p enhanced CoCl₂‐induced injury. In summary, an ischemia‐induced injury may be enhanced by a high level of NEAT1 through targeting miR‐27a‐3p. Thus, NEAT1 has the potential to be explored as a biomarker for diagnosis and target for therapeutic strategies in ischemia‐induced AKI.