T-Synthase Deficiency Enhances Oncogenic Features in Human Colorectal Cancer Cells via Activation of Epithelial-Mesenchymal Transition.
T-Synthase Deficiency Enhances Oncogenic Features in Human Colorectal Cancer Cells via Activation of Epithelial-Mesenchymal Transition.
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T-合酶缺乏通过激活上皮-间质转化增强人类结直肠癌细胞的致癌特征
DOI：
10.1155/2018/9532389
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发表时间：
2018
影响因子：
--
通讯作者：
Wen T
中科院分区：
文献类型：
--
作者：
Dong X;Jiang Y;Liu J;Liu Z;Gao T;An G;Wen T
背景未成熟的截短O-聚糖如Tn抗原在人结直肠癌（CRC）中经常被检测到;然而，Tn抗原表达对CRC的精确病理后果尚不清楚。T-合成酶是合成成熟O-聚糖的关键酶。在这里，我们研究了由T-合酶缺陷介导的结直肠癌细胞中Tn抗原表达的功能作用。方法为了敲除T合酶，我们使用CRISPR-Cas9技术靶向CRC细胞系（HCT 116）中编码T合酶的基因C1 GALT 1。通过Western印迹法证实T合酶的缺失，并通过流式细胞术测定HCT 116细胞中Tn抗原的表达。然后，我们评估了T-合酶缺乏对HCT 116细胞致癌行为的生物学效应。此外，我们通过确定上皮间质转化（EMT）途径分析了T-合酶缺陷在癌细胞中的机制作用。结果我们发现，强制敲除HCT 116细胞中的T合酶可显着诱导Tn抗原表达，这代表异常O-糖基化的发生。T-合酶的丧失显著增强了细胞的增殖和粘附，以及在培养中的迁移和侵袭力。更重要的是，我们证明了T-合酶缺乏直接诱导癌细胞的经典EMT特征。在T-合酶基因敲除的HCT 116细胞中，上皮细胞标志物E-钙粘蛋白的表达显著降低，同时间充质标志物包括snail和纤连蛋白（FN）的表达增强。结论大肠癌细胞T合酶缺陷不仅导致O-糖基化异常，还可能触发EMT通路的分子过程，从而增加肿瘤的侵袭和转移。
Background Immature truncated O-glycans such as Tn antigen are frequently detected in human colorectal cancer (CRC); however, the precise pathological consequences of Tn antigen expression on CRC are unknown. T-synthase is the key enzyme required for biosynthesis of mature O-glycans. Here we investigated the functional roles of Tn antigen expression mediated by T-synthase deficiency in CRC cells. Methods To knock out T-synthase, we used CRISPR-Cas9 technology to target C1GALT1, the gene encoding T-synthase, in a CRC cell line (HCT116). Deletion of T-synthase was confirmed by western blotting, and expression of Tn antigen was determined by flow cytometry in HCT116 cells. We then assessed the biological effects of T-synthase deficiency on oncogenic behaviors in HCT116 cells. Furthermore, we analyzed the mechanistic role of T-synthase deficiency in cancer cells by determining the epithelial-mesenchymal transition (EMT) pathway. Results We showed that forced knockout of T-synthase in HCT116 cells significantly induced Tn antigen expression, which represented the occurrence of aberrant O-glycosylation. Loss of T-synthase significantly enhanced cell proliferation and adhesion, as well as migration and invasiveness in culture. More importantly, we demonstrated that T-synthase deficiency directly induced classical EMT characteristics in cancer cells. E-cadherin, a typical epithelial cell marker, was markedly decreased in T-synthase knockout HCT 116 cells, accompanied by an enhanced expression of mesenchymal markers including snail and fibronectin (FN). Conclusions These findings indicate that T-synthase deficiency in CRC cells not only is responsible for aberrant O-glycosylation, but also triggers the molecular process of EMT pathway, which may translate to increased invasiveness and metastasis in cancers.
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影响因子：
9.7
作者：
HAMADA, S;FURUMOTO, H;AONO, T
通讯作者：
AONO, T
影响因子：
2.7
作者：
Sakai, Keiko;Yuasa, Noriyuki;Fujita-Yamaguchi, Yoko
通讯作者：
Fujita-Yamaguchi, Yoko
DOI：
10.1038/nrgastro.2013.120
发表时间：
2013-10
期刊：
Nature reviews. Gastroenterology & hepatology
影响因子：
--
作者：
通讯作者：
--
影响因子：
10.1
作者：
Scheid E;Major P;Bergeron A;Finn OJ;Salter RD;Eady R;Yassine-Diab B;Favre D;Peretz Y;Landry C;Hotte S;Mukherjee SD;Dekaban GA;Fink C;Foster PJ;Gaudet J;Gariepy J;Sekaly RP;Lacombe L;Fradet Y;Foley R
通讯作者：
Foley R
影响因子：
5.6
作者：
Khalili AA;Ahmad MR
通讯作者：
Ahmad MR