Gradiz Rui, Silva Henriqueta C, Carvalho Lina, Botelho Maria Filomena, Mota-Pinto Anabela
General Pathology Laboratory, Faculty of Medicine, University of Coimbra, Coimbra, Portugal.
CIMAGO - Research Center for Environment, Genetics and Oncobiology, Faculty of Medicine, University of Coimbra, Coimbra, Portugal.
Sci Rep. 2016 Feb 17;6:21648. doi: 10.1038/srep21648.
Studies using cell lines should always characterize these cells to ensure that the results are not distorted by unexpected morphological or genetic changes possibly due to culture time or passage number. Thus, the aim of this study was to describe those MIA PaCa-2 and PANC-1 cell line phenotype and genotype characteristics that may play a crucial role in pancreatic cancer therapeutic assays, namely neuroendocrine chemotherapy and peptide receptor radionuclide therapy. Epithelial, mesenchymal, endocrine and stem cell marker characterization was performed by immunohistochemistry and flow cytometry, and genotyping by PCR, gene sequencing and capillary electrophoresis. MIA PaCa-2 (polymorphism) expresses CK5.6, AE1/AE3, E-cadherin, vimentin, chromogranin A, synaptophysin, SSTR2 and NTR1 but not CD56. PANC-1 (pleomorphism) expresses CK5.6, MNF-116, vimentin, chromogranin A, CD56 and SSTR2 but not E-cadherin, synaptophysin or NTR1. MIA PaCA-1 is CD24(-), CD44(+/++), CD326(-/+) and CD133/1(-), while PANC-1 is CD24(-/+), CD44(+), CD326(-/+) and CD133/1(-). Both cell lines have KRAS and TP53 mutations and homozygous deletions including the first 3 exons of CDKN2A/p16(INK4A), but no SMAD4/DPC4 mutations or microsatellite instability. Both have neuroendocrine differentiation and SSTR2 receptors, precisely the features making them suitable for the therapies we propose to assay in future studies.
使用细胞系的研究应始终对这些细胞进行特征描述,以确保结果不会因可能由于培养时间或传代次数导致的意外形态或基因变化而失真。因此,本研究的目的是描述那些可能在胰腺癌治疗试验(即神经内分泌化疗和肽受体放射性核素治疗)中起关键作用的MIA PaCa-2和PANC-1细胞系的表型和基因型特征。通过免疫组织化学和流式细胞术进行上皮、间质、内分泌和干细胞标志物特征分析,并通过PCR、基因测序和毛细管电泳进行基因分型。MIA PaCa-2(多态性)表达CK5.6、AE1/AE3、E-钙黏蛋白、波形蛋白、嗜铬粒蛋白A、突触素、SSTR2和NTR1,但不表达CD56。PANC-1(多形性)表达CK5.6、MNF-116、波形蛋白、嗜铬粒蛋白A、CD56和SSTR2,但不表达E-钙黏蛋白、突触素或NTR1。MIA PaCA-1是CD24(-)、CD44(+/++)、CD326(-/+)和CD133/1(-),而PANC-1是CD24(-/+)、CD44(+)、CD326(-/+)和CD133/1(-)。两种细胞系均有KRAS和TP53突变以及包括CDKN2A/p16(INK4A)前3个外显子的纯合缺失,但无SMAD4/DPC4突变或微卫星不稳定性。两者均有神经内分泌分化和SSTR2受体,正是这些特征使其适合我们计划在未来研究中进行检测的治疗方法。