The ENO1 gene encodes a multifunctional enzyme that has been identified as a key component of the glycolytic pathway. ENO1 overexpression and post-translational modifications could be of diagnostic and prognostic value in many cancer types. The results of the ENO1 expression profiling of ovarian follicles suggest that ENO1 may play an important dual role in the progress of follicular development, where ENO1 acts as a glycolytic enzyme and also mediates apoptosis. ENO1 overexpression could make the primary culture follicle granulosa cells in vitro improve progesterone secretion. The over-expression of ENO1 protein can enhance the abilities of proliferation and migration in gastric cancer cells of AGS, which indicates that ENO1 may be an important potential tumor-marker associated with the development of gastric cancer. ENO1 and GPI can be used as markers of human sperm freezability before starting the cryopreservation procedure. The inhibition of ENO1 expression may be a novel strategy for therapy for NHLs patients, and it may be a target for drug resistance.