Objective To investigate the regulatory mechanism of long-chain non-coding RNA prostate cancer- related transcription 1 (PCAT1) in proliferation, migration, invasion and glutamine decomposition of tongue squamous cell carcinoma (TSCC) cells. Methods Real-time quantitative polymerase chain reaction was used to detect expression of PCAT1 and miR-329-3p in TSCC tissues, adjacent tissues, TSCC cells and normal oral keratinocytes. Liposomal transfection was applied to UM1 cells. Groups were the pcDNA group ( transfected with pcDNA), pcDNA-PCAT1 group (transfected with pcDNA-PCAT1), miR-NC group (transfected with miR-NC), miR-329-3p group (transfected with miR- 329-3p mimics), pcDNA-PCAT1 + miR-NC group (cotransfected with pcDNA-PCAT1 and miR-NC) and pcDNA-PCAT1 + miR-329-3p group ( cotransfected with pcDNA-PCAT1 and miR-329-3p mimics). Cell counting kit, migration assay (Transwell) and enzyme-linked immunosorbent assay were used to assess cell proliferation, migration / invasion, and levels of glutamine decomposition products glutamate and α-ketoglutarate (α-KG). The dual luciferase assay was used to detect binding between PCAT1 and miR-329-3p in cells. Results Compared with adjacent tissue, expression of PCAT1 in cancer tissue was upregulated significantly and expression of miR-329-3p was downregulated significantly. Compared with NHOK cells, PCAT1 expression was upregulated and miR-329-3p expression was downregulated in UM1, CAL-27 and SCC25 cells and of (P< 0. 05). Compared with the pcDNA group, expression of PCAT1 was significantly increased in the pcDNA- PCAT1 group, cell proliferation, invasion and migration were upregulated, and the levels of glutamate and α-KG were increased significantly (P<0. 05). PCAT1 targeted miR-329-3p. Compared with the miR-NC group, expression of miR- 329-3p in TSCC cells of the miR-329-3p group was increased significantly cell proliferation, migration, and invasion were reduced significantly, and the levels of glutamate and α-KG were reduced significantly ( P< 0. 05). Compared with the pcDNA-PCAT1+miR-NC group, expression of PCAT1 in the pcDNA-PCAT1+miR-329-3p group was reduced significantly, cell proliferation, migration, and invasion were reduced significantly, and the levels of glutamate and α-KG were also reduced significantly (P< 0. 05). Conclusions PCAT1 promotes the proliferation, migration, invasion, and glutamine decomposition of TSCC cells and its mechanism is related to targeting miR-329-3p.