Inhibitory effect of microRNA-154 targeting WHSC1 on cell proliferation of human skin squamous cell carcinoma through mediating the P53 signaling pathway.

Abstract

Skin squamous cell carcinoma (SCC) is a common, morbid, and frequently lethal malignancy and ranks as the sixth most deadly cancer worldwide. Hence, this study aims to explore the effect of microRNA-154 (miR-154) targeting WHSC1 on proliferation and apoptosis of SCC cells via the P53 signaling pathway. The targeting relationship between WHSC1 and miR-154 was validated using dual-luciferase reporter assay. Normal human epidermal keratinocytes (NHEK) were included, and SCC A431 and SCC-15 cell lines were cultured and transfected with miR-154 mimic, miR-154 inhibitor or siRNA-WHSC1. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) and western blot analysis were used for the miR-154 expression and levels of WHSC1, P53 signaling pathway- and apoptosis-related genes. MTT assay and flow cytometry were applied to determine the cell viability and apoptosis. WHSC1 is a target gene of miR-154. MiR-154 negatively regulated WHSC1 expression and inhibited the activation of P53 signaling pathway. In response to miR-154 mimic or siRNA-WHSC1, SCC A431 and SCC-15 cell lines exhibited increased expression of P73, P16 and Bax, decreased expression of WHSC1, P53, c-myc and Bcl-2, as well as attenuated cell viability and enhanced cell apoptosis. The treatment of miR-154 inhibitor reversed the tendency. These results demonstrate that up-regulation of miR-154 inhibits proliferation and induces apoptosis of human skin SCC cells by down-regulating WHSC1 and blocking the P53 signaling pathway.