Cancer-associated mutations reveal a novel role for EpCAM as an inhibitor of cathepsin-L and tumor cell invasion.
Cancer-associated mutations reveal a novel role for EpCAM as an inhibitor of cathepsin-L and tumor cell invasion.
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DOI：
10.1186/s12885-021-08239-z
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发表时间：
2021-05-12
期刊：
影响因子：
3.8
通讯作者：
Gillanders WE
中科院分区：
文献类型：
--
作者：
Sankpal NV;Brown TC;Fleming TP;Herndon JM;Amaravati AA;Loynd AN;Gillanders WE
EpCAM（上皮细胞粘附分子）在上皮癌中经常失调。先前的研究表明EpCAM参与调节致癌信号通路和上皮-间质转化。最近证实EpCAM含有甲状腺球蛋白1型（TY-1）结构域。已知具有TY-1结构域的多种蛋白质抑制组织蛋白酶-L（CTSL），CTSL是促进肿瘤细胞侵袭和转移的半胱氨酸蛋白酶。对人类癌症测序研究的分析表明，高达5.1%的测试肿瘤中存在体细胞EpCAM突变。查询癌症体细胞突变目录（COSMIC）数据库，以将癌症相关EpCAM突变的位置和氨基酸变化制成表格。为了确定EpCAM突变如何影响癌症生物学，我们研究了C66 Y，这是一种在肝癌中发现的破坏性TY-1结构域突变，以及其他13种癌症相关的EpCAM突变。使用体外和体内模型来确定野生型（WT）和突变体EpCAM对CTSL活性和侵袭的影响。免疫沉淀和定位研究测试EpCAM和CTSL蛋白结合，并确定EpCAM突变体的区室表达模式。我们证明了WT EpCAM而不是C66 Y EpCAM在体外抑制CTSL活性，并且EpCAM的TY-1结构域负责这种抑制。WT EpCAM，而不是C66 Y EpCAM，在体外抑制肿瘤细胞侵袭和在体内抑制肺转移。在一组扩展的人类癌细胞系中，EpCAM表达与CTSL活性呈负相关。先前的研究已经证明EpCAM种系突变可以阻止EpCAM在细胞表面表达。我们证明了C66 Y和多种其他EpCAM癌症相关突变阻止EpCAM的表面表达。阻止EpCAM细胞表面表达的癌症相关突变消除了EpCAM抑制CTSL活性和肿瘤细胞侵袭的能力。这些研究揭示了EpCAM作为CTSL抑制剂的新作用，证实了多种癌症相关EpCAM突变的功能相关性，并表明携带EpCAM突变的癌症的治疗脆弱性。在线版本包含补充材料，可通过10.1186/s12885-021-08239-z获得。
EpCAM (Epithelial cell adhesion molecule) is often dysregulated in epithelial cancers. Prior studies implicate EpCAM in the regulation of oncogenic signaling pathways and epithelial-to-mesenchymal transition. It was recently demonstrated that EpCAM contains a thyroglobulin type-1 (TY-1) domain. Multiple proteins with TY-1 domains are known to inhibit cathepsin-L (CTSL), a cysteine protease that promotes tumor cell invasion and metastasis. Analysis of human cancer sequencing studies reveals that somatic EpCAM mutations are present in up to 5.1% of tested tumors. The Catalogue of Somatic Mutations in Cancer (COSMIC) database was queried to tabulate the position and amino acid changes of cancer associated EpCAM mutations. To determine how EpCAM mutations affect cancer biology we studied C66Y, a damaging TY-1 domain mutation identified in liver cancer, as well as 13 other cancer-associated EpCAM mutations. In vitro and in vivo models were used to determine the effect of wild type (WT) and mutant EpCAM on CTSL activity and invasion. Immunoprecipitation and localization studies tested EpCAM and CTSL protein binding and determined compartmental expression patterns of EpCAM mutants. We demonstrate that WT EpCAM, but not C66Y EpCAM, inhibits CTSL activity in vitro, and the TY-1 domain of EpCAM is responsible for this inhibition. WT EpCAM, but not C66Y EpCAM, inhibits tumor cell invasion in vitro and lung metastases in vivo. In an extended panel of human cancer cell lines, EpCAM expression is inversely correlated with CTSL activity. Previous studies have demonstrated that EpCAM germline mutations can prevent EpCAM from being expressed at the cell surface. We demonstrate that C66Y and multiple other EpCAM cancer-associated mutations prevent surface expression of EpCAM. Cancer-associated mutations that prevent EpCAM cell surface expression abrogate the ability of EpCAM to inhibit CTSL activity and tumor cell invasion. These studies reveal a novel role for EpCAM as a CTSL inhibitor, confirm the functional relevance of multiple cancer-associated EpCAM mutations, and suggest a therapeutic vulnerability in cancers harboring EpCAM mutations. The online version contains supplementary material available at 10.1186/s12885-021-08239-z.
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影响因子：
--
作者：
Dykes, Samantha S;Fasanya, Henrietta O;Siemann, Dietmar W
通讯作者：
Siemann, Dietmar W
影响因子：
3.8
作者：
Gostner JM;Fong D;Wrulich OA;Lehne F;Zitt M;Hermann M;Krobitsch S;Martowicz A;Gastl G;Spizzo G
通讯作者：
Spizzo G
DOI：
10.1111/j.1432-1033.1996.0125h.x
发表时间：
1996-08-15
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
Molina, F;Bouanani, M;Granier, C
通讯作者：
Granier, C
影响因子：
14.8
作者：
Blum, Galia;von Degenfeld, Georges;Bogyo, Matthew
通讯作者：
Bogyo, Matthew
影响因子：
2.9
作者：
Chen, Qingyong;Fei, Jun;Lu, Guohua
通讯作者：
Lu, Guohua