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Aims: This study aims to investigate the role of miR-144-3p and phosphatase and tensin homolog (PTEN) in nasopharyngeal carcinoma (NPC), along with their crosstalk with the phosphoinositide 3-kinase (PI3K)-protein kinase B (Akt) pathway., Methods: Quantitative reverse transcription polymerase chain reaction and western blot were used to measure the gene expression at the transcriptional and translational levels. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and colony formation assay were used to examine cell proliferation via standard protocol. Transwell assay was conducted to examine cell invasiveness. A flow cytometer was used to determine cell apoptosis. Dual-Luciferase Reporter Gene Assay (SLDL-100) was used to confirm the target relationship between miR-144-3p and PTEN. Xenografts were used to detect the in vivo effects of the molecules of interest., Results: miR-144-3p was significantly overexpressed, whereas PTEN was more underexpressed in tumor tissues than in adjacent tissues. miR-144-3p promoted the proliferation and invasion of NPC cells and inhibited apoptosis by directly targeting PTEN, which improves PI3K-Akt signaling. miR-144-3p forced epithelial-mesenchymal transition in NPC., Conclusion: miR-144-3p promotes the progression of NPC by directly targeting PTEN via crosstalk with PI3K-Akt signaling., (© 2019 Wiley Periodicals, Inc.)