Disruption of 3D tissue integrity facilitates adenovirus infection by deregulating the coxsackievirus and adenovirus receptor
Disruption of 3D tissue integrity facilitates adenovirus infection by deregulating the coxsackievirus and adenovirus receptor
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DOI：
10.1073/pnas.0337599100
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发表时间：
2003-02-18
影响因子：
11.1
通讯作者：
Korn, WM
中科院分区：
文献类型：
--
作者：
Anders, M;Hansen, R;Korn, WM
人柯萨奇病毒和腺病毒受体（CAR）代表了感染期间腺病毒附着的主要细胞位点。了解调节其表达的机制有助于提高基于腺病毒的治疗的有效性和安全性。我们在人乳腺癌进展的3D细胞培养模型中表征了CAR表达的调控，该模型模拟了体外生理组织背景的各个方面。表型正常的乳腺上皮细胞（S1）和它们的恶性衍生物（T4-2细胞）在组织培养塑料（2D）或3D培养物的基膜基质中生长。在3D中生长的S1细胞显示出低水平的CAR，其主要在细胞-细胞连接处表达。相反，T4-2细胞表达高水平的CAR，其主要在细胞质中。当通过表皮生长因子受体的信号传导在T4-2细胞中被抑制时，细胞恢复正常表型，CAR蛋白表达显著降低，并且蛋白质重新定位于细胞-细胞连接处。S1细胞作为2D培养物或在胶原蛋白-1（这些细胞的营养生理学微环境）中以3D生长，导致CAR上调至与T4-2细胞中的水平相似的水平，与细胞生长速率无关。因此，CAR的表达取决于上皮细胞的3D组织的完整性和极性。通过微环境的变化（包括恶性转化）破坏这种组织，导致CAR的上调，从而增强细胞对腺病毒感染的易感性。
The human coxsackievirus and adenovirus receptor (CAR) represents the primary cellular site of adenovirus attachment during infection. An understanding of the mechanisms regulating its expression could contribute to improving efficacy and safety of adenovirus-based therapies. We characterized regulation of CAR expression in a 3D cell culture model of human breast cancer progression, which mimics aspects of the physiological tissue context in vitro. Phenotypically normal breast epithelial cells (S1) and their malignant derivative (T4-2 cells) were grown either on tissue culture plastic (2D) or 3D cultures in basement membrane matrix. S1 cells grown in 3D showed low levels of CAR, which was expressed mainly at cell-cell junctions. In contrast, T4-2 cells expressed high levels of CAR, which was mainly in the cytoplasm. When signaling through the epidermal growth factor receptor was inhibited in T4-2 cells, cells reverted to a normal phenotype, CAR protein expression was significantly reduced, and the protein relocalized to cell-cell junctions. Growth of S1 cells as 2D cultures or in 3D in collagen-l, a noriphysiological microenvironment for these cells, led to up-regulation of CAR to levels similar to those in T4-2 cells, independently of cellular growth rates. Thus, expression of CAR depends on the integrity and polarity of the 3D organization of epithelial cells. Disruption of this organization by changes in the microenvironment, including malignant transformation, leads to up-regulation of CAR, thus enhancing the cell's susceptibility to adenovirus infection.