X-H Wei1, X-L Gu, X-T Zhou, M Ma, C-X Lou. 1. Department of Dermatology, the Third Hospital of Ji'nan, Ji'nan, China. Wei717@163.com.
Abstract
OBJECTIVE: To explore the possible role and mechanism of miR-497 in cutaneous squamous cell carcinoma. PATIENTS AND METHODS: Quantitative Real-time polymerase chain reaction (qRT-PCR) was used to detect miR-497 and FAM114A2 expression level in 38 cases of cutaneous squamous cell carcinoma (CSCC) and 22 normal skin tissues as well as in CSCC cell lines (A431, HSC-5) and normal cells (HaCaT). MiR-497 effects on cell proliferation and cell cycle were examined by CCK8 assays and flow cytometry. Dual luciferase reporter gene assay was performed to detect the regulating relationship between miR-497 and FAM114A2. In addition, the expression of FAM114A2 after overexpression or knockdown of miR-497 was detected by Western blot to evaluate whether miR-497 could regulate proliferation and cell cycle by regulating the expression of FAM114A2. RESULTS: MiR-497mRNA expression in CSCC tissues and cell lines was markedly lower than that in normal tissues and cells. Meanwhile, FAM114A2 mRNA and protein levels in CSCC tissues were markedly higher when compared to than that in normal tissues. miR-497 overexpression or knockdown could inhibit or promote the cell proliferation and cell cycle of A431, HSC-5. The dual luciferase reporter gene assay suggested that FAM114A2 might be a direct target gene of miR-497, and that FAM114A2 expression had a significant negative correlation with miR-497. Overexpression of miR-497 could inhibit FAM114A2 protein expression. Besides, FAM114A2 knockdown reversed the inhibitory effect of low expression of miR-497 on proliferation rate of A431 or HSC-5 cells. CONCLUSIONS: MiR-497 was lowly expressed in squamous cell carcinoma tissues and cells, which can participate in the regulation of cell proliferation through FAM114A2, thus promoting the progression of CSCC.
OBJECTIVE: To explore the possible role and mechanism of miR-497 in cutaneous squamous cell carcinoma. PATIENTS AND METHODS: Quantitative Real-time polymerase chain reaction (qRT-PCR) was used to detect miR-497 and FAM114A2 expression level in 38 cases of cutaneous squamous cell carcinoma (CSCC) and 22 normal skin tissues as well as in CSCC cell lines (A431, HSC-5) and normal cells (HaCaT). MiR-497 effects on cell proliferation and cell cycle were examined by CCK8 assays and flow cytometry. Dual luciferase reporter gene assay was performed to detect the regulating relationship between miR-497 and FAM114A2. In addition, the expression of FAM114A2 after overexpression or knockdown of miR-497 was detected by Western blot to evaluate whether miR-497 could regulate proliferation and cell cycle by regulating the expression of FAM114A2. RESULTS: MiR-497mRNA expression in CSCC tissues and cell lines was markedly lower than that in normal tissues and cells. Meanwhile, FAM114A2 mRNA and protein levels in CSCC tissues were markedly higher when compared to than that in normal tissues. miR-497 overexpression or knockdown could inhibit or promote the cell proliferation and cell cycle of A431, HSC-5. The dual luciferase reporter gene assay suggested that FAM114A2 might be a direct target gene of miR-497, and that FAM114A2 expression had a significant negative correlation with miR-497. Overexpression of miR-497 could inhibit FAM114A2 protein expression. Besides, FAM114A2 knockdown reversed the inhibitory effect of low expression of miR-497 on proliferation rate of A431 or HSC-5 cells. CONCLUSIONS:MiR-497 was lowly expressed in squamous cell carcinoma tissues and cells, which can participate in the regulation of cell proliferation through FAM114A2, thus promoting the progression of CSCC.