Purpose of investigation: microRNAs (miRNAs), which can regulate cell biological behaviors such as proliferation and apoptosis as oncogenes or anti-oncogenes, are closely associated with cancer onset and progression. The aim of this study was to detect the expression changes of miR-101 in cervical cancer tissues and the effects on the biological functions of cervical cancer SiHa cells. Materials and Methods: Through transient transfection of SiHa cells with mature miR-101 sequences, the effects on apoptosis, proliferation, and cell cycle were evaluated by real-time PCR, CCK-8 assay, and flow cytometry. Results: Significantly less miR-101 was expressed in cervical cancer tissues than that in normal cervical tissues. miR-101 was overexpressed in SiHa cells through transient transfection of miR-101 mimics. CCK-8 assay and flow cytometry showed that miR-101 overexpression significantly inhibited cell proliferation, promoted apoptosis, and arrested them in the Gj/S phase. Real-time PCR exhibited that Mcl-1 and c-Fos mRNA expressions significantly decreased. Conclusion: miR-101 significantly reduced the viability of SiHa cells as a potential anti-oncogene.