Huan Yang1, Chunhua Shen. 1. Department of Dermatology, Affiliated Wujiang Hospital of Nantong University, Suzhou City, China.
Abstract
PURPOSE: Melanoma is a malignant skin tumor that can easily metastasize, while no effective treatment exists for this disease. This study explored the mechanism of microRNA-29c in inhibiting melanoma cell growth. METHODS: Bioinformatics analysis and polymerase chain reaction (PCR) experiments were performed to analyze the expression of microRNA-29c in various samples. The Cell Counting Kit-8 (CCK-8) experiment was used to detect cell viability. The mimic and inhibitor of microRNA-29c were transfected into melanoma cells to achieve microRNA-29c overexpression or knockdown so as to observe the biological effect on the melanoma cells. Flow cytometry was used to detect cell cycle, while the luciferase reporter gene assay was used for predicting microRNA-29c target genes. Western blot was performed to determine the cellular protein expression. RESULTS: microRNA-29c was highly expressed in melanoma cells. Overexpression of microRNA-29c inhibited cell viability and induced G1 cell cycle arrest. Conversely, cell proliferation and cycle progression were promoted by transfection of microRNA-29c inhibitor in melanoma cells. In addition, CDK6 served as a microRNA-29c target gene. G1 phase of melanoma cells was blocked by knockdown of CDK6. CONCLUSIONS: microRNA-29c can inhibit the growth of melanoma cells by targeting CDK6, which could trigger G1 arrest of melanoma cells.
PURPOSE:Melanoma is a malignant skin tumor that can easily metastasize, while no effective treatment exists for this disease. This study explored the mechanism of microRNA-29c in inhibiting melanoma cell growth. METHODS: Bioinformatics analysis and polymerase chain reaction (PCR) experiments were performed to analyze the expression of microRNA-29c in various samples. The Cell Counting Kit-8 (CCK-8) experiment was used to detect cell viability. The mimic and inhibitor of microRNA-29c were transfected into melanoma cells to achieve microRNA-29c overexpression or knockdown so as to observe the biological effect on the melanoma cells. Flow cytometry was used to detect cell cycle, while the luciferase reporter gene assay was used for predicting microRNA-29c target genes. Western blot was performed to determine the cellular protein expression. RESULTS:microRNA-29c was highly expressed in melanoma cells. Overexpression of microRNA-29c inhibited cell viability and induced G1 cell cycle arrest. Conversely, cell proliferation and cycle progression were promoted by transfection of microRNA-29c inhibitor in melanoma cells. In addition, CDK6 served as a microRNA-29c target gene. G1 phase of melanoma cells was blocked by knockdown of CDK6. CONCLUSIONS:microRNA-29c can inhibit the growth of melanoma cells by targeting CDK6, which could trigger G1 arrest of melanoma cells.
Authors: Yulia Andreevna Veryaskina; Sergei Evgenievich Titov; Igor Fyodorovich Zhimulev Journal: Med Princ Pract Date: 2022-03-30 Impact factor: 2.132