Novel organotypic culture model of cholangiocarcinoma progression.
Novel organotypic culture model of cholangiocarcinoma progression.
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DOI：
10.1111/j.1872-034x.2012.01026.x
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发表时间：
2012-11
期刊：
影响因子：
--
通讯作者：
Sirica AE
中科院分区：
文献类型：
--
作者：
Campbell DJ;Dumur CI;Lamour NF;Dewitt JL;Sirica AE
最近的研究表明，促结缔组织增生间质中α-平滑肌肌动蛋白阳性肌纤维母细胞（α-SMA阳性CAF）的增加可能与肝内胆管癌（ICC）患者的更具侵袭性的癌症和更差的生存结局有关。为了便于研究与ICC进展相关的α-SMA阳性CAFs和胆管癌细胞之间的细胞和分子相互作用，我们开发了一种新的胆管癌三维（3-D）器官型培养模型，该模型更准确地模拟了体内促结缔组织增生性ICC的基质微环境，基因表达谱和选择的病理生理学特征。通过在I型胶原凝胶基质中共培养一株胆管癌细胞（来源于胆管接种自发转化的大鼠胆管细胞后在同系大鼠肝脏中形成的ICC）与不同数量的克隆α-SMA阳性CAF（从相同肿瘤类型建立），建立了该独特模型。单一培养的胆管癌细胞和α-SMA阳性CAF均表现出基质成肌纤维细胞与恶性胆管细胞类型的细胞特异性生物标志物基因表达谱。相比之下，器官型共培养所表现出的基因表达谱和组织病理学特征与亲本原位ICC的全组织样品的基因表达谱和组织病理学特征非常相似。我们进一步显示α-SMA阳性CAF在体外显著增强胆管癌细胞“导管样”生长和癌细胞迁移/侵袭，以及促进已知与ICC侵袭相关的选定基因的上调表达。这种新的器官型模型为研究微环境对胆管癌体外进展的影响提供了重要的新资源，并可能作为临床前模型用于鉴定分子靶向治疗。
Recent studies have suggested that increased α-smooth muscle-actin-positive myofibroblastic cells (α-SMA-positive CAFs) in the desmoplastic stroma may relate to a more aggressive cancer and worse survival outcomes for intrahepatic cholangiocarcinoma (ICC) patients. To facilitate investigating cellular and molecular interactions between α-SMA-positive CAFs and cholangiocarcinoma cells related to ICC progression, we developed a novel 3-dimensional (3-D) organotypic culture model of cholangiocarcinoma that more accurately mimics the stromal microenvironment, gene expression profile, and select pathophysiological characteristics of desmoplastic ICC in vivo. This unique model was established by co-culturing within a type I collagen gel matrix, a strain of cholangiocarcinoma cells (derived from an ICC formed in syngeneic rat liver following bile duct inoculation of spontaneously-transformed rat cholangiocytes) with varying numbers of clonal α-SMA-positive CAFs established from the same tumor type. Cholangiocarcinoma cells and α-SMA-positive CAFs in monoculture each exhibited cell specific biomarker gene expression profiles characteristic of stromal myofibroblastic cell versus malignant cholangiocyte cell types. In comparison, the gene expression profile and histopathological characteristics exhibited by the organotypic co-culture closely resembled those of whole tissue samples of the parent orthotopic ICC. We further showed α-SMA-positive CAFs to significantly enhance cholangiocarcinoma cell “ductal-like” growth and cancer cell migration/invasiveness in vitro, as well as to promote up-regulated expression of select genes known to be associated with ICC invasion. This novel organotypic model provides an important new resource for studying the effects of microenvironment on cholangiocarcinoma progression in vitro and may have potential as a preclinical model for identifying molecularly targeted therapies.