15 December 2019 : Laboratory Research
A Positive Tetraspanin 8 (TSPAN8)/β-Catenin Regulatory Loop Enhances the Stemness of Colorectal Cancer CellsZhengyu Zhan1BCDE, Luxing Zhong1DE, Miao Feng1DEF, Yuling Guo1ABG*
Med Sci Monit 2019; 25:9594-9601
BACKGROUND: The expression of TSPAN8 (tetraspanin 8) is upregulated in colorectal cancer; however, its roles in colorectal cancer progression are never been revealed. This work aimed to investigate TSPAN8 effects and the molecular basis in regulating colorectal cancer stemness.
MATERIAL AND METHODS: Real-time quantitative polymerase chain reaction and western blot analysis were used to detect the expression of TSPAN8 expression in clinical samples and the expression of stemness genes in colorectal cancer cells. Sphere forming analysis was performed to detect TSPAN8 effects on sphere forming ability of colorectal cancer cells. Co-IP and ChIP analysis were performed to confirm the molecular basis contributing to TSPAN8-mediated effects on colorectal cancer stemness.
RESULTS: TSPAN8 expression is increased in colorectal cancer tissues. Knockdown of TSPAN8 reduced the expression of stemness genes and sphere forming capacity in colorectal cancer cells. Mechanistically, TSPAN8 directly interacted β-catenin and enhanced its protein expression, which is necessary for TSPAN8-mediated effects on colorectal cancer stemness. Conversely, β-catenin directly bound to TSPAN8 promoter and enhanced TSPAN8 transcription.
CONCLUSIONS: TSPAN8 promotes colorectal cancer stemness through a positive TSPAN8/β-catenin regulatory loop.
Keywords: Colorectal Neoplasms, Neoplastic Stem Cells, Wnt Signaling Pathway, Disease Progression, primary cell culture, Tetraspanins, transcriptome, beta Catenin
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