Colorectal cancer (CRC) is a prevalent malignancy worldwide, and microRNAs (miRNAs) have been recognized for their significant role in CRC progression and potential as therapeutic targets. This study aimed to investigate the impact of miR-222-3p on CRC cell proliferation, migration, and invasion, along with its target genes. HT29 cells were transfected with mimic-negative control (mimic-NC) or mimic-miR-222-3p, while the control group remained untreated. Cell proliferation, migration, and invasion were assessed using CCK-8 and Transwell assays. qRT-PCR and Western blotting were employed to measure gene mRNA and protein expression, respectively. A luciferase reporter assay verified the binding between miR-222-3p and its downstream target gene ITGB3. The results revealed that enhanced miR-222-3p expression significantly increased HT29 cell proliferation, migration, and invasion. qRT-PCR and Western blotting indicated reduced expression of ITGB3 and E-cadherin, and upregulation of Vimentin and α-SMA by miR-222-3p. The luciferase reporter assay confirmed ITGB3 as a direct target of miR-222-3p. In conclusion, miR-222-3p promotes CRC progression by regulating ITGB3 expression, suggesting its potential as a crucial biomarker and therapeutic target for colorectal cancer.