The aim of this study is to unveil the role of integrin in influencing the differentiation of PC12 cell grown on PLLA aligned nanofibers by integrative study of cDNA microarray, proteomics technology and microRNA sequencing technology. First, PLLA-aligned nanofibers were prepared by electrospinning, and the chemical composition and surface morphology of the nanofibers were examined. Then, morphology and neurite length of PC12 cells and GRGDS-treated PC12 cells on PLLA-aligned nanofibers (AF and GA group) were measured by high-content analysis system. Subsequently, protein expression profile was analyzed by iTRAQ proteomics technology and 250 differentially expressed proteins were screened in the GA group. Integrative analysis of mRNAmicroRNA-protein data showed that 'MAPK signaling pathway' was the key affected pathway after the function of integrin were interfered by GRGDS. Other seven pathways were also influenced, and the differentiation of PC12 cell grown on PLLA aligned nanofibers was finally inhibited.