In this paper, the cDNA microarray technique was used to investigate the gene expression profiles of human fibroblasts irradiated by low-intensity red light. Proliferation assays showed that the fibroblast HS27 cells responded with a curve effect to different doses of low-intensity red light irradiation at a wavelength of 628 nm. An optimal dose of 0.88 J per cm2 was chosen for subsequent cDNA microarray experiments. The gene expression profiles revealed that 111 genes were regulated by the red light irradiation and can be grouped into 10 functional categories. Most of these genes directly or indirectly play roles in the enhancement of cell proliferation and the suppression of apoptosis. Two signaling pathways, the p38 mitogen-activated protein kinase signaling pathway and the platelet-derived growth factor signaling pathway, were found to be involved in cell growth induced by irradiation of low-intensity red light. Several genes related to antioxidation and mitochondria energy metabolism were also found to express differentially upon irradiation. This study provides insight into the molecular mechanisms associated with the beneficial effects of red light irradiation in accelerating wound healing.