Anethole suppresses the growth of human skin cancer cells by targeting microRNA498/STAT4 axis

Abstract

Purpose: To investigate the effects of anethole on human skin cancer cells. Methods: Cell viability was determined using CCK-8 and colony formation assay, while AO/EB and Annexin V/PI assays were used to assess apoptosis. The mRNA expressions of genes were assayed by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Results: The growth of skin cancer cells was significantly reduced by anethole (p < 0.05). On the other hand, normal human skin cells were marginally affected by anethole. Furthermore, anethole significantly suppressed the colony formation potential of CRL-6475 skin cancer cells in a concentration-dependent manner (p < 0.05). Results from AO/EB and annexin V/PI staining assays revealed that anethole induced apoptosis in skin cancer cells, while qRT-PCR results indicate that anethole significantly up-regulated microRNA-498 in skin cancer cells (p < 0.05). However, anethole had no effect on the expression levels of miR-7, miR-9, miR-145, miR-31, miR-27, miR-498 miR-1298, miR-1299, 1179, miR-375, miR-508 and miR-23. CCK-8 assay results confirmed that miR-498 up-regulation significantly mimicked the anti-proliferative effects of anethole. Bioinformatics analysis and luciferase reporter assay led to identification of STAT4 as the regulatory target of miR-498. Furthermore, silencing of STAT4 significantly mimicked the effects of anethole administration (p < 0.05). Conclusion: These results indicate that anethole inhibits skin cancer cell growth by targeting the miR-498/STAT4 axis, and therefore, has potentials for use in the management of skin cancer.