SAFB通过调节Wnt信号通路活性促进结直肠癌增殖

SAFB promotes colorectal cancer proliferation by regulating Wnt signaling pathway activity

目的探讨SAFB在结直肠癌增殖过程中的作用及相关分子机制。方法采用免疫组化、Western blot和RT-PCR检测结直肠癌石蜡组织和新鲜配对组织中SAFB的表达,分析SAFB表达与肿瘤分期的关系;采用MTT、平板克隆及裸鼠皮下成瘤等实验检测SAFB对结直肠癌细胞增殖能力的影响;利用公共数据库(GEO数据库等),通过基因富集分析(gene set enrichment analysis,GSEA)检测SAFB基因表达相关信号通路的富集情况;采用双荧光素酶报告基因实验、Western blot法及免疫荧光实验检测Wnt信号通路的活性。结果Oncomine及TCGA在线数据库分析结果显示,结直肠癌组织中SAFB表达水平降低(P<0.01),且随着Dukes分期的增高,表达水平明显下降(P<0.05)。MTT、平板克隆及裸鼠皮下成瘤实验结果显示,SAFB高表达在体内外均能显著抑制肿瘤细胞的增殖。GSEA及双荧光素酶报告基因实验结果显示,SAFB低表达与Wnt通路激活密切相关。结论SAFB在结直肠癌中低表达,并与Wnt信号通路激活密切相关,可能通过Wnt信号通路调节肿瘤细胞的增殖能力。

Purpose To investigate the role and molecular mechanism of SAFB in the proliferation of colorectal cancer(CRC).Methods Immunohistochemistry,Western blot and RT-PCR were used to detect the expression of SAFB in CRC paraffin-embedded samples and fresh paired tissue samples,and to analyze the relationship between SAFB expression and tumor stage.MTT,plate cloning,soft AGAR colony formation and subcutaneous tumor formation in nude mice were used to detect the effects of SAFB on the proliferation of colorectal cancer cells.The enrichment of SAFB gene expression related signaling pathways was detected by GSEA using public databases(GEO database,etc.).Double luciferase assay,Western blot and immunofluorescence assay were used to detect the activity of Wnt signaling pathway.Results Oncomine and TCGA online database analysis showed that SAFB expression decreased in CRC tissues(P<0.01),and decreased significantly with the increase of Dukes stage.MTT assay,plate cloning,soft AGAR colony assay and subcutaneous tumor-forming experiment in nude mice showed that high expression of SAFB could significantly inhibit the proliferation of tumor cells in vitro and in vivo.GSEA and dual luciferase reporter assay demonstrated that low SAFB expression was closely related to Wnt pathway activation.Conclusion SAFB is low expressed in colorectal cancer and is closely related to the activation of Wnt signaling pathway,which may regulate the proliferation of tumor cells through the Wnt signaling pathway.