miR-627-5p inhibits malignant progression of cervical cancer by targeting ANGPTL4

Abstract

In recent years, accumulating evidence has highlighted the critical role of miR-627-5p in the occurrence and progression of various cancers. However, its specific role and mechanism in cervical cancer (CC) remain unclear. This study aimed to elucidate the mechanism by which miR-627-5p inhibits the malignant progression of CC and assess its potential clinical implications. In C33A cells, the mRNA expression levels of ANGPTL4 and miR-627-5p were analyzed using qRT-PCR. The miR-627-5p mimics and their control (miR-NC) were transfected into C33A cells to determine whether miR-627-5p directly regulates ANGPTL4 expression. A comprehensive suite of assays, including CCK-8, migration, transwell, flow cytometry, and Western blotting, was conducted to evaluate how miR-627-5p modulates the malignant biological behavior of CC cells. Rescue experiments were performed by overexpressing ANGPTL4. In C33A cells, miR-627-5p expression was reduced, whereas ANGPTL4 expression was elevated. Further analysis confirmed that miR-627-5p negatively regulates ANGPTL4 by directly targeting its 3'-UTR. Functional assays demonstrated that miR-627-5p inhibits proliferation, invasion, migration, and epithelial-mesenchymal transition (EMT) while promoting apoptosis and S-phase arrest in C33A cells, effects that were reversed by ANGPTL4 overexpression. These findings highlight the potential of miR-627-5p as both a biomarker and a therapeutic target for CC. By inhibiting EMT and regulating ANGPTL4 expression, miR-627-5p may provide a novel avenue for improving therapeutic strategies, particularly in advanced or metastatic CC. Moreover, miRNA-based therapies, supported by advanced delivery systems such as nanoparticle carriers, could enhance the stability and precision of miR-627-5p applications. This study lays the groundwork for future research integrating miR-627-5p into precision medicine approaches for CC treatment.

Figure 1.

Expression and interaction of miR-627-5p and ANGPTL4 in C33A cells. A,B) qRT-PCR was used to detect the mRNA levels of miR-627-5p and ANGPTL4 in H8 and C33A cells, showing lower miR-627-5p expression and higher ANGPTL4 expression in C33A cells compared to H8 cells. C) DLR assays and bioinformatic analysis confirmed that miR-627-5p directly targets ANGPTL4 through binding to its 3'-UTR. D-F) qRT-PCR and WB were performed after transfection with miR-627-5p mimics. The results indicated that miR-627-5p mimics upregulated miR-627-5p expression while downregulating ANGPTL4 mRNA and protein levels, confirming their negative regulatory relationship in C33A cells. **p<0.01; ***p<0.001.

Figure 2.

MiR-627-5p impact on C33A cell malignant evolvement. A-D) CCK-8, clonal formation, wound-healing, and Transwell assays were used to evaluate the effects of miR-627-5p mimics on C33A cell proliferation, migration, and invasion. The results showed that miR-627-5p inhibited proliferation, slowed wound healing, and reduced invasive ability in C33A cells; scale bars (50 μm) were added to all relevant images, and representative images were chosen based on reproducible outcomes across at least three independent experiments. E) WB analysis demonstrated that miR-627-5p mimics upregulated E-cadherin protein expression while downregulating N-cadherin and vimentin levels, indicating inhibition of EMT. *p<0.05; **p<0.01; ***p<0.001.

Figure 3.

MiR-627-5p impact on C33A cycle and apoptosis. A,B) Flow cytometer and WB were used to assess apoptosis rates and apoptosis-related protein expression in C33A cells transfected with miR-627-5p mimics. miR-627-5p promoted apoptosis by upregulating Bax and downregulating Bcl-2 protein levels. C) Flow cytometer analysis revealed that miR-627-5p induced S-phase arrest, suggesting its role in disrupting cell cycle progression. *p<0.05; **p<0.01; ***p<0.001.

Figure 4.

MiR-627-5p controls ANGPTL4 to inhibit the growth of C33A cells. A) qRT-PCR analysis confirmed the successful overexpression of ANGPTL4 in C33A cells transfected with oe-ANGPTL4. B-E) Functional assays, including CCK-8, clonal formation, wound-healing, and Transwell assays, demonstrated that ANGPTL4 overexpression counteracted the inhibitory effects of miR-627-5p mimics on cell proliferation, migration, and invasion; microscopy images now include scale bars for improved precision; selected images represent typical outcomes observed across three independent experiments, ensuring reproducibility; scale bars: 50 μm. F) Western blot analysis showed that ANGPTL4 overexpression restored EMT-related protein levels, reversing the effects of miR-627-5p mimics. *p<0.05; **p<0.01; ***p<0.001.

Figure 5.

MiR-627-5p promotes apoptosis and S phase arrest in C33A cells by regulating ANGPTL4. A,B) Flow cytometer and WB showed that ANGPTL4 overexpression reduced apoptosis and S-phase arrest, counteracting the effects of miR-627-5p mimics; ANGPTL4 overexpression increased Bcl-2 levels while decreasing Bax levels, reducing apoptosis. C) Flow cytometer confirmed that ANGPTL4 overexpression alleviated the S-phase arrest induced by miR-627-5p, further highlighting their regulatory relationship in cervical cancer cells. *p<0.05; **p<0.01; ***p<0.001.