Non-small cell lung cancer (NSCLC) is demonstrated as one of the most common malignant tumors and accounts for about 25% of cancer-related deaths each year. Extensive bodies of studies have manifested that microRNAs (miRNAs) play pivotal roles in the development of numerous malignant tumors by involving in modulation of cell biological processes. Although miR-4319 has been validated to execute tumor suppressor properties in triple-negative breast cancer, explorations on the function and latent mechanism of miR-4319 participating in NSCLC are still unclear. In this study, we proved that miR-4319 acted as a tumor suppressor in NSCLC progression via restraining cell proliferation and migration as well as boosting apoptosis. Further, miR-4319 bound with LIN28 and negatively regulated the expression of LIN28. Our data unveiled that LIN28 promoted RFX5 mRNA stability and miR-4319 led to the destabilization of RFX5 by targeting LIN28. In addition, RFX5 motivated the transcription of YAP and enhanced expression of YAP abolished the miR-4319 upregulation-mediated suppressive regulation of NSCLC tumorigenesis. In conclusion, miR-4319 dampened YAP expression to mitigate the tumorigenesis of NSCLC through inhibiting LIN28-mediated RFX5 stability, which offered an insight into the molecular mechanism underlying miR-4319 in NSCLC development.