Effects of Apigenin on Expressions of MiR-342-3p and its Target Gene FOXM1 and Self-Renewal in Cervical Cancer HeLa Cells

ZHAO Xia; LE Yijie; CAO Jianguo; LIU Jie; DONG Weilei

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Journal of Hunan Normal University(Medical Science) ›› 2024, Vol. 21 ›› Issue (6) : 22-28.

Effects of Apigenin on Expressions of MiR-342-3p and its Target Gene FOXM1 and Self-Renewal in Cervical Cancer HeLa Cells

ZHAO Xia¹, LE Yijie², CAO Jianguo², LIU Jie¹, DONG Weilei¹

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Abstract

Objective To investigate whether apigenin (API) inhibits self-renewal by downregulating FOXM1 expression via miR-342-3p upregulation in cervical cancer HeLa cells. Methods HeLa cells transfected with miR-342-3p inhibitor or FOXM1 cDNA were treated with sub-cytotoxicity concentrations of API (1 μM, 3 μM and 10 μM) or in combination with API (10 μM) treatment and miR-342-3p inhibitor or FOXM1 cDNA. Sphere formation assay was used to detect the self-renewal in HeLa cells. Quantitative real-time RT-PCR (qRT-PCR) was employed to detect miRNAs and mRNA expression levels. Immunoblotting was utilized to detect the expression levels of FoxM1, CD49f and Sox2 protein. The luciferase assay was applied to examine if FOXM1 is directly targeted by miR-342-3p. The efficacy of API on tumor growth in HeLa-derived sphere-forming cells (SFCs) in vivo was determined in a nude mouse xenograft model. Results The subcellular concentration of API (1 μM; 3 μM and 10 μM) decreased sphere formation rate [(32.70±2.48)% vs.(23.67±2.14)%, (16.50±1.66)%, (11.52±1.05)%, all of P<0.05)], down-regulated CD49f and Sox2 protein levels as well as elevated miR-342-3p levels and reduced FoxM1 protein expression levels in HeLa cells, in a dose-dependent tendency. Moreover, miR-342-3p inhibitor or FOXM1 cDNA could antagonize the aforementioned efficacy of API. In addition, oral API (50 and 100mg/kg) administration resulted in markedly reduced tumor growth [tumor weight: (0.66±0.11)g vs.(0.33±0.03)g, (0.17±0.02)g, all of P<0.05]. Luciferase assay demonstrated FOXM1 is directly targeted by miR-342-3p. Conclusion API reduces self-renewal capacity in cervical cancer HeLa cells by targeting FOXM1 via miR-342-3p upregulation.

Key words

HeLa cells / self-renewal / apigenin / miR-342-3p / FOXM1 / action mechanism

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ZHAO Xia, LE Yijie, CAO Jianguo, LIU Jie, DONG Weilei. Effects of Apigenin on Expressions of MiR-342-3p and its Target Gene FOXM1 and Self-Renewal in Cervical Cancer HeLa Cells[J]. Journal of Hunan Normal University(Medical Science). 2024, 21(6): 22-28

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