Gastric cancer (GC) is the fourth most common malignancy worldwide, with 80% mortality rate in over 70% countries. Recently, targeted therapy for GC has great clinical prospects, and it is still badly needed to find novel molecular targets to control the progression and development of GC. Kinesin family member 3B (KIF3B) is known as a microtubule motor kinesin and one of the most ubiquitously expressed KIFs. KIF3B participates in multiple cellular processes such as mitosis and spermatogenesis, and the possible role of KIF3B on tumor progression has been widely revealed. KIF3B affects the progression and metastasis of multiple types of tumors, such as pancreatic cancer, prostate cancer, and hepatocellular carcinoma; however, its potential impact on GC is still unknown. Herein, we explored the possible role of KIF3B on the progression of GC and noticed that KIF3B was high expression in tumor tissues from GC patients. KIF3B was also significantly correlated with clinical pathological characteristics such as tumor size (P = .014*) and recurrence (P = .044*). We further revealed that KIF3B depleted GC cells exhibited impaired proliferation capacity in vitro. Similarly, KIF3B depletion suppressed tumor growth of GC cells in mice. In conclusion, we identified KIF3B as a promising therapeutic target for the treatment of GC.