Potential therapeutic targets discovery by transcriptome analysis of an in vitro human gastric signet ring carcinoma model
Potential therapeutic targets discovery by transcriptome analysis of an in vitro human gastric signet ring carcinoma model
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DOI：
10.1007/s10120-022-01307-8
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发表时间：
2022-06
期刊：
影响因子：
7.4
通讯作者：
K. Yamaguchi;Tomoyasu Yoshihiro;H. Ariyama;Mamoru Ito;M. Nakano;Yuichiro Semba;Jumpei Nogami;K. Tsu
中科院分区：
文献类型：
--
作者：
K. Yamaguchi;Tomoyasu Yoshihiro;H. Ariyama;Mamoru Ito;M. Nakano;Yuichiro Semba;Jumpei Nogami;K. Tsu
背景E-钙粘素表达缺失在印戒癌中常见。编码E-cadherin的CDH 1基因发生种系突变的人，发生弥漫性胃癌的几率更高。E-cadherin表达的缺失被认为是引发肿瘤发生的重要因素。MethodsTo investigate new therapeutic targets for gastric SRCC，we engineered an E-cadherin-deficient SRCC model in vitro using a human gastric organoid（hGO）withCDH 1 knockout（KO）. ResultsCDH 1 KO hGO细胞表现出与SRCC相似的形态学变化和高细胞运动性。RNA测序显示，与野生型相比，CDH 1 KO hGO细胞中基质金属蛋白酶（MMP）基因上调。MMP抑制剂在体外抑制CDH 1 KO hGO细胞和SRCC细胞系的细胞运动。95例临床胃癌组织的免疫荧光分析显示，MMP-3在E-cadherin异常的SRCC中特异性丰富。另外，CDH 1 KO后CXCR 4分子转位到细胞膜上。在培养基中加入CXCR 4的配体CXCL 12可延长CDH 1 KO hGO细胞的存活时间，并被抑制剂AMD 3100所消除。在临床SRCC样品中，CXCL 12分泌成纤维细胞表现出显着的浸润到癌area.ConclusionsE-cadherin缺陷SRCCs可能获得细胞运动通过上调基质金属蛋白酶。CXCL 12阳性的癌症相关成纤维细胞可以作为一个小生境来维持癌细胞的存活。MMPs和CXCL 12/CXCR 4轴代表了作为E-cadherin缺陷SRCC的新治疗靶点的有希望的候选者。
BackgroundLoss of E-cadherin expression is frequently observed in signet ring carcinoma (SRCC). People with germline mutations inCDH1,which encodes E-cadherin, develop diffuse gastric cancer at a higher rate. Loss of E-cadherin expression is thus assumed to trigger oncogenic development.MethodsTo investigate novel therapeutic targets for gastric SRCC, we engineered an E-cadherin-deficient SRCC model in vitro using a human gastric organoid (hGO) withCDH1knockout (KO).ResultsCDH1KO hGO cells demonstrated distinctive morphological changes similar to SRCC and high cell motility. RNA-sequencing revealed up-regulation of matrix metalloproteinase (MMP) genes inCDH1KO hGO cells compared to wild type. MMP inhibitors suppressed cell motility ofCDH1KO hGO cells and SRCC cell lines in vitro. Immunofluorescent analysis with 95 clinical gastric cancer tissues revealed that MMP-3 was specifically abundant in E-cadherin-aberrant SRCC. In addition, CXCR4 molecules translocated onto the cell membrane afterCDH1KO. Addition of CXCL12, a ligand of CXCR4, to the culture medium prolonged cell survival ofCDH1KO hGO cells and was abolished by the inhibitor, AMD3100. In clinical SRCC samples, CXCL12-secreting fibroblasts showed marked infiltration into the cancer area.ConclusionsE-cadherin deficient SRCCs might gain cell motility through upregulation of MMPs. CXCL12-positive cancer-associated fibroblasts could serve to maintain cancer-cell survival as a niche. MMPs and the CXCL12/CXCR4 axis represent promising candidates as novel therapeutic targets for E-cadherin-deficient SRCC.