As one of the most deadly malignant tumours, lung cancer has been increasing in incidence and mortality worldwide. The incidence and mortality rates of lung cancer are increasing annually. A deeper understanding of the development of lung cancer at the molecular level is of great significance for accurate diagnosis and efficient treatment of lung cancer. The serine/arginine-rich splicing factors (SRSFs) family contains 12 highly homologous proteins, namely SRSF1-SRSF12, which have been reported to play an important role in the development of various tumours. Here, we found that the expression of multiple SRSF proteins was upregulated in lung cancer, and prognostic analyses revealed that only SRSF3 was associated with the prognosis of lung cancer patients. We found that the differentially expressed genes regulated by SRSF3 were heavily enriched in some tumour progression-related signalling pathways, such as p53, glycolysis signalling pathway, etc. We found that SRSF3 was also strongly associated with the expression of m6A-related genes, ferroptosis-related genes and some common immune checkpoints. We also found that SRSF3 differential expression in lung cancer patients was associated with sensitivity to some common drugs used to treat lung cancer. Interestingly, we also found that SRSF3 can undergo phase separation and promotes proliferation of lung cancer cells. These results suggest that SRSF3 may serve as a potential target for the diagnosis and treatment of lung cancer.