Dynamic Change of Polarity in Primary Cultured Spheroids of Human Colorectal Adenocarcinoma and Its Role in Metastasis
Dynamic Change of Polarity in Primary Cultured Spheroids of Human Colorectal Adenocarcinoma and Its Role in Metastasis
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DOI：
10.1016/j.ajpath.2015.12.011
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发表时间：
2016-04-01
影响因子：
6
通讯作者：
Inoue, Masahiro
中科院分区：
文献类型：
--
作者：
Okuyama, Hiroaki;Kondo, Jumpei;Inoue, Masahiro
肠上皮细胞具有顶基极性，控制着营养物质和废物的交换。细胞极性的扰动似乎是癌症的普遍特征，尽管大多数结直肠癌是分化腺癌，其中极性在一定程度上保持。人们对极性失调在癌症中的作用知之甚少。采用癌组织起源球体法制备和培养球体。球状体在悬浮液或I型胶原凝胶中培养。极性通过免疫组化和电镜观察。在类球体中观察到两种极性状态：一种是尖向内，在I型胶原凝胶中，类球体的顶膜位于腔内；顶向外，顶膜位于悬浮球体的最外层。这些极性是高度可互换的。Src和dynamin抑制剂减弱了极性转换。在患者中，侵袭血管的癌细胞簇具有尖向内和尖向外的形态特征，而原发性和转移性肿瘤具有尖向内的特征。在小鼠肝转移模型中，注入门静脉的尖向外球体在几天内变成肝脏的尖向内球体。Src和dynamin抑制剂显著降低肝转移。在球状体和人类癌症中观察到极性转换。极性转换在实验性肝转移模型中是至关重要的。
Intestinal epithelial cells possess apical-basal polarity, which governs the exchange of nutrients and waste. Perturbation of cell polarity appears to be a general feature of cancers, although most colorectal cancers are differentiated adenocarcinomas, in which polarity is maintained to some extent. Little is known about the role of dysregulated polarity in cancer. The cancer tissue originated spheroid method was applied to the preparation and culture of spheroids. Spheroids were cultured in suspension or in type I collagen gel. Polarity was assessed by IHC of apical markers and electron microscopy. Two types of polarity status in spheroids were observed: apical-in, with apical membrane located at cavities inside the spheroids in type I collagen gel; and apical-out, with apical membrane located at the outermost layer of spheroids in suspension. These polarities were highly interchangeable. Inhibitors of Src and dynamin attenuated the polarity switch. In patients, clusters of cancer cells that invaded vessels had both apical-in and apical-out morphologic features, whereas primary and metastatic tumors had apical in features. In a mouse Liver metastasis model, apical-out spheroids injected into the portal vein became apical-in spheroids in the liver within a few days. Inhibitors of Src and dynamin significantly decreased liver metastasis. Polarity switching was observed in spheroids and human cancer. The polarity switch was critical in an experimental liver metastasis model.