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Objective To clarify the polo-like kinase1 ( PLK1) expression in human gastric cancer tissue and its clinicopathological significance in gastric carcinoma. Methods PLK1 expression in 60 cancer tissues and their corresponding noncancerous tissues from gastric cancer patients was measured by both real-time quantitative RT-PCR and western blot assay. Immunohistochemistry was used to detect PLK1 protein expression in eighty-nine paraffin-embedded samples. Results The PLK1 mRNA and protein expression level in the 60 fresh cancer tissues was significantly higher than that in noncancerous tissues ( P ＜0. 0001, P =0. 031 respectively). In paraffin-embed-ded samples, apart from its increased expression level, PLK1 was found to be in both cytoplasm and nucleus, double-site location only occurred in poor-differentiated cancer, PLK1 expression intensity was associated with tumor differentiation ( P = 0. 03 ), invasion ( P = 0. 032 ), TNM stage ( P = 0. 019 ) , ki67 expression ( P = 0. 011 ). The patients with negative PLK1 expression had better survival rate than that with positive PLKI expression ( P = 0. 0292 ).Conclusion PLK1 may have clinicopathological value in tumor diagnosis, and may become another new biomarker and therapeutic target for gastric cancer.