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五种通过体外培养方法建立的人胰腺导管腺癌细胞系。

Five primary human pancreatic adenocarcinoma cell lines established by the outgrowth method.

机构信息

Department of Visceral, Thoracic, and Vascular Surgery, University Hospital Carl Gustav Carus, Technische Universität Dresden, Germany.

出版信息

J Surg Res. 2012 Jan;172(1):29-39. doi: 10.1016/j.jss.2011.04.021. Epub 2011 May 7.

Abstract

BACKGROUND

Pancreatic ductal adenocarcinoma is an aggressive tumor; treatment remains a challenge because of the lack of effective therapeutic strategies. Basic research in this field is dependent on the availability of model systems. New pancreatic cancer cell lines are therefore important for the study of its biology. In the present study, we report the establishment and characterization of five new pancreatic cancer cell lines (PaCaDD-43, -60, -119, -135, -137).

MATERIAL AND METHODS

All cell lines were derived from pancreatic ductal adenocarcinomas by the Dresden outgrowth protocol. The five cell lines originated from primary pancreatic tumors, lymph node metastases, or malignant pleural effusions. We characterized the cell lines by examining their morphology and their cytostructural and functional profiles.

RESULTS

All cell lines grew as adherent monolayers and were cultured in optimized Dresden-medium. The doubling time ranged from 22 to 47 h. v-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog (KRAS) mutations were detected in four of the five cell lines. KRAS mutations were identical between each primary tumor and the cell line derived from it. Immunohistochemical staining showed cytoplasmic expression of CK8/18, mostly membrane and partially cytoplasmic expression of E-cadherin and strong expression of ezrin in all cell lines. Three cell lines showed nuclear p53 accumulation and heterogeneous expression of vimentin. SMAD4 was heterogeneously expressed in four of the cell lines.

CONCLUSIONS

We were able to establish five new primary pancreatic carcinoma cell lines. As applicable tools for basic research, these cell lines might contribute to a better understanding and treatment of this aggressive tumor.

摘要

背景

胰腺导管腺癌是一种侵袭性肿瘤;由于缺乏有效的治疗策略,治疗仍然是一个挑战。该领域的基础研究依赖于模型系统的可用性。因此,新的胰腺癌细胞系对于研究其生物学具有重要意义。本研究报告了 5 种新的胰腺癌细胞系(PaCaDD-43、-60、-119、-135、-137)的建立和鉴定。

材料与方法

所有细胞系均通过德累斯顿体外培养法从胰腺导管腺癌中分离得到。这 5 种细胞系来源于原发肿瘤、淋巴结转移或恶性胸腔积液。我们通过观察细胞形态、细胞结构和功能特征对细胞系进行了鉴定。

结果

所有细胞系均作为贴壁单层生长,并在优化的德累斯顿培养基中培养。倍增时间为 22-47 小时。在 5 种细胞系中的 4 种中检测到 v-Ki-ras2 Kirsten 大鼠肉瘤病毒癌基因同源物(KRAS)突变。每个原发肿瘤与其衍生的细胞系之间的 KRAS 突变是相同的。免疫组化染色显示 CK8/18 细胞质表达,E-钙粘蛋白主要为膜表达,部分为细胞质表达,ezrin 在所有细胞系中表达强烈。3 种细胞系显示核 p53 积累和波形蛋白异质性表达。SMAD4 在 4 种细胞系中呈异质性表达。

结论

我们成功建立了 5 种新的原发性胰腺癌细胞系。作为基础研究的适用工具,这些细胞系可能有助于更好地理解和治疗这种侵袭性肿瘤。

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