PDF(Pubmed)
Abstract：
SLC2A5 is a high-affinity fructose transporter, which is frequently upregulated in multiple human malignant tumours. However, the function and molecular mechanism of SLC2A5 in colorectal cancer (CRC) remain unknown.
We detected the expression levels of SLC2A5 in CRC tissues and CRC cell lines by western blotting, qRT-PCR and immunohistochemistry. CRC cell lines with stable overexpression or knockdown of SLC2A5 were constructed to evaluate the functional roles of SLC2A5 in vitro through conventional assays. An intrasplenic inoculation model was established in mice to investigate the effect of SLC2A5 in promoting metastasis in vivo. Methylation mass spectrometry sequencing, methylation specific PCR, bisulphite sequencing PCR, ChIP-qPCR and luciferase reporter assay were performed to investigate the molecular mechanism underlying transcriptional activation of SLC2A5.
We found that SLC2A5 was upregulated in colorectal tumour tissues. Functionally, a high level of SLC2A5 expression was associated with increased invasion and metastasis capacities of CRC cells both in vitro and in vivo. Mechanistically, we unveiled that S100P could integrate to a specific region of SLC2A5 promoter, thereby reducing its methylation levels and activating SLC2A5 transcription.
Our results reveal a novel mechanism that S100P mediates the promoter demethylation and transcription activation of SLC2A5, thereby promoting the metastasis of CRC.
We detected the expression levels of SLC2A5 in CRC tissues and CRC cell lines by western blotting, qRT-PCR and immunohistochemistry. CRC cell lines with stable overexpression or knockdown of SLC2A5 were constructed to evaluate the functional roles of SLC2A5 in vitro through conventional assays. An intrasplenic inoculation model was established in mice to investigate the effect of SLC2A5 in promoting metastasis in vivo. Methylation mass spectrometry sequencing, methylation specific PCR, bisulphite sequencing PCR, ChIP-qPCR and luciferase reporter assay were performed to investigate the molecular mechanism underlying transcriptional activation of SLC2A5.
We found that SLC2A5 was upregulated in colorectal tumour tissues. Functionally, a high level of SLC2A5 expression was associated with increased invasion and metastasis capacities of CRC cells both in vitro and in vivo. Mechanistically, we unveiled that S100P could integrate to a specific region of SLC2A5 promoter, thereby reducing its methylation levels and activating SLC2A5 transcription.
Our results reveal a novel mechanism that S100P mediates the promoter demethylation and transcription activation of SLC2A5, thereby promoting the metastasis of CRC.
摘要：
SLC2A5是一种高亲和力果糖转运蛋白，在多种人类恶性肿瘤中经常上调。然而,SLC2A5在结直肠癌（CRC）中的功能和分子机制尚不清楚。
我们通过蛋白质印迹法检测了SLC2A5在CRC组织和CRC细胞系中的表达水平，qRT-PCR和免疫组织化学。构建具有SLC2A5的稳定过表达或敲低的CRC细胞系以通过常规测定在体外评估SLC2A5的功能作用。在小鼠中建立了脾内接种模型，以研究SLC2A5在体内促进转移的作用。甲基化质谱测序，甲基化特异性PCR，亚硫酸氢盐测序PCR，进行ChIP-qPCR和荧光素酶报告基因测定以研究SLC2A5转录激活的分子机制。
我们发现SLC2A5在结直肠肿瘤组织中上调。功能上,SLC2A5的高水平表达与CRC细胞在体外和体内的侵袭和转移能力增加相关.机械上,我们揭示了S100P可以整合到SLC2A5启动子的特定区域，从而降低其甲基化水平并激活SLC2A5转录。
我们的结果揭示了S100P介导SLC2A5启动子去甲基化和转录激活从而促进CRC转移的新机制。
我们通过蛋白质印迹法检测了SLC2A5在CRC组织和CRC细胞系中的表达水平，qRT-PCR和免疫组织化学。构建具有SLC2A5的稳定过表达或敲低的CRC细胞系以通过常规测定在体外评估SLC2A5的功能作用。在小鼠中建立了脾内接种模型，以研究SLC2A5在体内促进转移的作用。甲基化质谱测序，甲基化特异性PCR，亚硫酸氢盐测序PCR，进行ChIP-qPCR和荧光素酶报告基因测定以研究SLC2A5转录激活的分子机制。
我们发现SLC2A5在结直肠肿瘤组织中上调。功能上,SLC2A5的高水平表达与CRC细胞在体外和体内的侵袭和转移能力增加相关.机械上,我们揭示了S100P可以整合到SLC2A5启动子的特定区域，从而降低其甲基化水平并激活SLC2A5转录。
我们的结果揭示了S100P介导SLC2A5启动子去甲基化和转录激活从而促进CRC转移的新机制。
