Long-term primary culture of a clear cell ovarian carcinoma reveals an epithelial-mesenchymal cooperative interaction.
Long-term primary culture of a clear cell ovarian carcinoma reveals an epithelial-mesenchymal cooperative interaction.
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DOI：
10.1186/s12935-015-0243-8
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发表时间：
2015
影响因子：
5.8
通讯作者：
Telleria CM
中科院分区：
文献类型：
--
作者：
Goyeneche AA;Koch M;Bell MC;Telleria CM
我们研究了从卵巢透明细胞癌（O-CCC）活检中获得的原代培养物，通过（a）评估其保留肿瘤起源的体外病理学特征的能力;（B）表征从复杂肿块中释放的主要细胞，而无需强制纯化任何特定的细胞实体;（c）研究其长期增殖能力。从诊断为O-CCC的盆腔肿块中培养原代细胞。通过相差显微镜进行细胞培养物的形态学分析。上皮细胞、间充质细胞和肿瘤起始细胞的标记物通过免疫细胞化学进行评价。通过检测掺入新合成DNA的溴脱氧尿苷（BrdU）来研究细胞增殖。作为O-CCC的生物标志物，我们评估了肝细胞核因子（HNF）1β的表达。我们发现，上皮细胞的形态特征表达E-钙粘蛋白，并扩大随着时间的推移，在文化中，事实上，BrdU的掺入证实。然而，表达间充质标记波形蛋白的具有间充质样特征的细胞分配到上皮隔室的边缘。此外，我们还发现一些上皮细胞也表达波形蛋白。在孵育开始时，超过60%的原代细胞表达O-CCC标志物HNF 1 β;该百分比在传代后下降。我们发现，上皮细胞而不是间充质细胞进行DNA复制，并且上皮细胞和间充质细胞中很少有细胞表达干细胞样肿瘤抗原CD 133。我们提供了从O-CCC活检中大量分离的细胞可以在培养中维持数月的原理证明，并且两个一致的细胞区室-一个保留O-CCC标记物HNF 1 β的上皮细胞和另一个间充质细胞-持续存在，并且似乎具有合作的相互作用，导致上皮细胞在间充质细胞环境中增殖。
We studied a primary culture developed from a biopsy of a clear cell carcinoma of the ovary (O-CCC) by (a) assessing its capacity to retain in vitro pathological features of the tumor of origin; (b) characterizing the main cells released from the complex mass without forced purification of any particular cellular entity; and (c) investigating its long-term proliferative capacity. A primary cell culture was developed from a pelvic mass diagnosed as an O-CCC. The morphological analysis of the cell culture was carried out by phase contrast microscopy. Markers of epithelial, mesenchymal, and tumor initiating cells were evaluated by immunocytochemistry. Cell proliferation was studied by detection of bromodeoxyuridine (BrdU) incorporated into newly synthesized DNA. As a biomarker of O-CCC, we assessed the expression of hepatocyte nuclear factor (HNF) 1β. We show that cells with epithelial morphological features express E-cadherin and expand with time in culture, a fact that the incorporation of BrdU confirms. Cells with mesenchymal-like characteristics that express the mesenchymal marker vimentin, however, allocate to the edges of the epithelial compartment. Moreover, we found that some cells with epithelial features also expressed vimentin. At the beginning of incubation, over 60 % of primary cells expressed the O-CCC marker HNF1β; such percentage declined upon passaging. We show that epithelial not mesenchymal cells undergo DNA replication, and that few cells in both epithelial and mesenchymal compartments express the stem-like tumor antigen CD133. We provide proof-of-principle that cells separated in bulk from a biopsy of an O-CCC can be maintained in culture for several months, and that two consistent cellular compartments—one epithelial that retains the O-CCC marker HNF1β, and another mesenchymal—persist, and seem to have a cooperative interaction leading to the multiplication of epithelial cells within a mesenchymal cellular environment.