Tacrolimus (FK506) is widely used to prevent rejection of transplanted organs following organ transplantation. However, the cytotoxicity of FK506 in human T lymphocytes has not been fully studied. MicroRNAs (miRNA) are well-studied small noncoding RNAs that regulate the expression of genes at the posttranscriptional level. In this study, we found that tacrolimus suppresses glucose metabolism through the induction of miR-202. Moreover, the expression levels of miR-202 and hexokinase 2 exhibited a reverse correlation under FK506 treatments: FK506 induced miR-202 but suppressed hexokinase 2. Both microRNA target prediction and western blotting results showed that hexokinase 2 is a target of miR-202 in Jurkat cells. We found that the overexpression of miR-202 inhibited hexokinase 2 expression, resulting in decreased glucose consumption. Importantly, inhibition of miR- 202 rendered Jurkat cells sensitive to tacrolimus through the promotion of glucose demand. Our study will provide new insights into the microRNA-mediated immunotherapy of cancer cells.