Polo-like kinase 1 (Plk1) is an interesting molecule both as a biomarker and as a target for highly specific cancer therapy for several reasons. However, the functional significance of Plk1 in renal cell carcinoma (RCC) has not been reported. To explore whether Plk1 plays a general role in renal carcinoma, we examined the expression of Plk1 protein in renal urothelial carcinoma and cell lines, and analyzed the relationship between Plk1 protein expression and development, proliferation, and invasion of renal carcinoma. Immunohistochemisty was used to detect the expression of Plk1 in 100 renal carcinoma tissues. Moreover, the expression of Plk1 was analyzed by western blot and real-time polymerase chain reaction (PCR) in 80 renal carcinoma tissues and 20 normal renal tissues. CCK-8 assay, colony formation assay, and Transwell assay were used to examine proliferation and invasion ability of renal cancer cells with treatment of scytonemin (the specific inhibitor of Plk1). Statistical analysis was used to discuss the association between Plk1 expression and clinicopathologic parameters, and proliferation and invasion ability of renal cancer cells. Plk1 expressions were greater in cancerous tissues than in normal tissues (P<0.05). With an increase in tumor grade and stage, tumor metastasis, and recurrence, the level of Plk1 increased significantly in renal cancerous tissues. Moreover, there was a significantly higher expression of Plk1 in higher degree of malignant renal adenocarcinoma cell ACHN than that in renal adenocarcinoma cell 769-P. With increasing concentration of scytonemin, we found that cell proliferation and invasion activity decreased significantly. Plk1 expression status was closely correlated with important histopathologic characteristics (grades, stages, metastasis, and recurrence) of renal carcinomas. Furthermore, Plk1 played an important function on renal cancer cells' proliferation and invasion.