In our study, we found that HR-8348 cells with down-regulated miRNA-96-5p was significantly reduced in their proliferation, migration, and invasion abilities compared with the untransfected HR-8348 cells, while GPC3 expression was significantly up-regulated in the transfected cells, which indirectly suggested that GPC3 might play a nonaggressive phenotype effect in rectal cancer cells. This evidence concerns the gene GPC3 and rectal cancer.