Drought stress significantly affects the growth, development, and yield of cotton, triggering the response of multiple genes. Among them, ascorbate peroxidase (APX) is one of the important antioxidant enzymes in the metabolism of reactive oxygen species in plants, and APX enhances the ability of plants to resist oxidation, thus increasing plant stress tolerance. Therefore, enhancing the activity of APX in cells is crucial to improving plant stress resistance. Previous studies have isolated differentially expressed proteins under drought stress (GhAPX7) in drought-resistant (KK1543) and drought-sensitive (XLZ26) plants. Thus, this study analyzed the expression patterns of GhAPX7 in different cotton tissues to verify the drought resistance function of GhAPX7 and explore its regulatory pathways. GhAPX7 had the highest expression in cotton leaves, which significantly increased under drought stress, suggesting that GhAPX7 is essential for improving antioxidant capacity and enzyme activities in cotton. GhAPX7 silencing indirectly affects pronounced leaf yellowing and wilting in drought-resistant and drought-sensitive plants under drought stress. Malondialdehyde (MDA) content was significantly increased and chlorophyll and proline content and APX enzyme activity were generally decreased in silenced plants compared to the control. This result indicates that GhAPX7 may improve drought resistance by influencing the contents of MDA, chlorophyll, proline, and APX enzyme activity through increased expression levels. Transcriptome analysis revealed that the drought-related differentially expressed genes between the control and treated groups enriched plant hormone signal transduction, MAPK signaling, and plant-pathogen interaction pathways. Therefore, the decreased expression of GhAPX7 significantly affects the expression levels of genes in these three pathways, reducing drought resistance in plants. This study provides insights into the molecular mechanisms of GhAPX7 and its role in drought resistance and lays a foundation for further research on the molecular mechanisms of response to drought stress in cotton.