Objective To observe the expression of long-chain non-coding RNA (lncRNA) metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) and hsa-miR155-3p in patients with rheumatoid arthritis (RA) and their relationship with Notch signaling pathway, and to explore the possible pathogenesis of RA. Methods Peripheral blood of RA patients (RA group) and healthy controls (NC group) were used to screen differentially expressed lncRNA and mRNA by high-throughput gene sequencing. Reverse-transcription PCR was used to detect the expression of lncRNA MALAT1, hsa-miR155-3p and Notch signaling pathway receptor ligands. Results The lncRNA sequencing analysis showed a total of 9158 differentially expressed lncRNAs. Gene ontology (GO) functional classification annotations revealed that the differentially expressed mRNA was mainly involved in immune inflammatory response, cellular transcriptional regulation and so on. Pathway analysis proved that differentially expressed mRNA was significantly related to the genes involved in rheumatoid arthritis, cellular senescence, and Notch signaling pathways. According to cis and trans prediction, Jagged1-2, Delta1-2 and Notch1-2 might be closely related to RA. MALAT1 in the RA group was lower than that in the NC group, and hsa-miR155-3p expression was significantly higher than that in the NC group. The expression of Notch pathway ligands Delta1, Delta2 Jagged1, Jagged2 and the receptors Notch1 and Notch2 in the RA group increased. Correlation analysis showed that hsa-miR155-3p was inversely proportional to MALAT1, hsa-miR155-3p was directly proportional to Notch pathway Delta1, Jagged1, and MALAT1 was inversely proportional to Jagged2. Conclusion In patients with rheumatoid arthritis, lncRNA MALAT1 is reduced and hsa-mir155-3p is raised, which jointly regulate the change of Notch signaling pathway.