When it comes to cervical cancer, it is the most common malignancy in gynecology. This study aimed to investigate the concomitant status of miRNA-9-5p in cervical cancer and explore its potential mechanism for treating cervical cancer. The levels of miRNA-9-5p, CA125, CA199, and CEA expression were detected by RT-PCR, and the downstream target genes regulated by miRNA-9-5p were screened by the Venn map. Cytoscape was utilized to find the binding sites of the two genes, and luciferase reporter assay verified the direct regulation of miRNA-9-5p and CXCR4; the CCK-8 assay detected its regulation on cell proliferation, and the expression of miRNA-9-5p, CXCR4, PCNA, Ki67 mRNA, and proteins was detected by RT-PCR and western blot. The expression of miRNA-9-5p was decreased, while the levels of CA125, CA199, and CEA were increased in the model group. The database predicts that CXCR4 is a gene regulated by miRNA-9-5p. The luciferase reporter gene results indicated that miRNA-9-5p could directly regulate the expression of CXCR4 and miRNAs are detected by intracellular transfer inhibitors. In total, MiRNA-9-5p can be utilized as a biological marker for cervical cancer that may inhibit cancer cells’ proliferation by inhibiting the expression of the CXCR4 gene and protein.