<p>The PI3K-AKT signaling pathway (SP) has been introduced as a key regulatory pathway in cervical cancer (CC). Inhibition of this SP could be a therapeutic strategy in CC. Our previous bioinformatics analysis exhibited that miR-542 could have a dual inhibitory function on this SP by targeting <i>PIK3CB</i> and <i>AKT1</i> genes with its two arms (-3p and -5p) and proposed it as an appropriate therapeutic target for CC. The current study experimentally investigated the dual inhibitory function of miR-542 on the PI3K-AKT SP. qRT-PCR was performed following transfection of the recombinant pEGFP-C1 vector containing miR-542 precursor into the CaSki and miR-542 overexpression to quantify the expression level of target genes of miR-542 (<i>AKT1</i> and <i>PIK3CB</i>) as regulators of PI3K-AKT SP and their downstream genes affecting cell proliferation and apoptosis (<i>CDKN1A</i> and <i>BCL2</i>). In addition, the effect of overexpression of miR-542 on cell cycle and apoptosis was examined by flow cytometry using propidium iodide and PE Annexin V/7-AAD staining, respectively. The recombinant cells showed a significant decrease in the expression of <i>AKT1</i>, <i>PIK3CB</i>, and <i>BCL2</i> genes, and a significant increase in the level of <i>CDKN1A</i> gene expression, simultaneously with the highest overexpression of miR-542 at 48 h post-transfection. Furthermore, the apoptosis was remarkably induced and the cell cycle was arrested in recombinant cells compared to mock cells. miR-542 promoted apoptosis and cell cycle arrest by dual inhibiting the PI3K/AKT SP. It may be introduced as an appropriate target for CC treatment.</p> Graphical Abstract <p></p>