Longitudinal Claudin Gene Expression Analyses in Canine Mammary Tissues and Thereof Derived Primary Cultures and Cell Lines

Hammer, Susanne Conradine; Becker, Annegret; Rateitschak, Katja; Mohr, Annika; Ripoli, Florenza Lüder; Hennecke, Silvia; Junginger, Johannes; Hewicker‐Trautwein, M.; Brenig, Bertram; Ngezahayo, Anaclet; Nolte, Ingo; Escobar, Hugo Murua

doi:10.3390/ijms17101655

Cited by 7 publications

(6 citation statements)

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“…Ki-67 was evaluated by the avidin-biotin complex technique as previously described 131 . No animal was killed for the generation of positive controls, all tissue specimens used as positive controls for this study were sent to the Department of Pathology for diagnosis.…”

Section: Immunohistochemical Examination the Expression Of Er Pr Hmentioning

confidence: 99%

Analysis of Copy-Number Variations and Feline Mammary Carcinoma Survival

Granados-Soler

Bornemann-Kolatzki²,

Beck³

et al. 2020

Sci Rep

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Feline mammary carcinomas (FMCs) are highly malignant. As the disease-free survival (DFS) and overall survival (OS) are short, prognostication is crucial. Copy-number variations (CNVs) analysis by nextgeneration sequencing serves to identify critical cancer-related genomic regions. Thirty-three female cats with FMCs were followed during two years after surgery. Tumours represented tubulopapillary and solid carcinomas encompassing six molecular subtypes. Regardless of the histopathological diagnosis, molecular subtypes showed important differences in survival. Luminal A tumours exhibited the highest DFS (p = 0.002) and cancer-specific OS (p = 0.001), and the lowest amount of CNVs (p = 0.0001). In contrast, basal-like triple-negative FMCs had the worst outcome (DFS, p < 0.0001; and OS, p < 0.00001) and were the most aberrant (p = 0.05). In the multivariate analysis, copy-number losses (CNLs) in chromosome B1 (1-23 Mb) harbouring several tumour-repressors (e.g. CSMD1, MTUS1, MSR1, DBC2, and TUSC3) negatively influenced DFS. Whereas, copy-number gains (CNGs) in B4 (1-29 Mb) and F2 (64-82.3 Mb) comprising epithelial to mesenchymal transition genes and metastasis-promoting transcription factors (e.g. GATA3, VIM, ZEB1, and MYC) negatively influenced DFS and cancer-specific OS. These data evidence an association between specific CNVs in chromosomes B1, B4 and F2, and poor prognosis in FMCs.

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Section: Immunohistochemical Examination the Expression Of Er Pr Hmentioning

confidence: 99%

Analysis of Copy-Number Variations and Feline Mammary Carcinoma Survival

Granados-Soler

Bornemann-Kolatzki²,

Beck³

et al. 2020

Sci Rep

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“…Immunophenotyping by the avidin-biotin method was performed in the original tumour and cell pellets of TiHo-0906 (low [P7], and high [P80] passages) as described elsewhere 46 using the following markers (details see Table 6 ): E-cad, CK8/18, pan-CK, CK14, CK5/6, p63, SMA, CALP, Vim, HMGA2, CD44, ER, PR, HER-2, COX-2, p53, CLDN-2 and proliferation marker Ki-67. Positive and negative controls used for immunohistochemistry are listed in the Supplementary Table 1 .…”

Section: Methodsmentioning

confidence: 99%

TiHo-0906: a new feline mammary cancer cell line with molecular, morphological, and immunocytological characteristics of epithelial to mesenchymal transition

Granados-Soler

Junginger

Hewicker‐Trautwein

et al. 2018

Sci Rep

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Feline mammary carcinomas (FMCs) with anaplastic and malignant spindle cells histologically resemble the human metaplastic breast carcinoma (hMBC), spindle-cell subtype. hMBCs display epithelial-to-mesenchymal transition (EMT) characteristics. Herein we report the establishment and characterization of a cell line (TiHoCMglAdcar0906; TiHo-0906) exhibiting EMT-like properties derived from an FMC with anaplastic and malignant spindle cells. Copy-number variations (CNVs) by next-generation sequencing and immunohistochemical characteristics of the cell line and the tumour were compared. The absolute qPCR expression of EMT-related markers HMGA2 and CD44 was determined. The growth, migration, and sensitivity to doxorubicin were assessed. TiHo-0906 CNVs affect several genomic regions harbouring known EMT-, breast cancer-, and hMBCs-associated genes as AKT1, GATA3, CCND2, CDK4, ZEB1, KRAS, HMGA2, ESRP1, MTDH, YWHAZ, and MYC. Most of them were located in amplified regions of feline chromosomes (FCAs) B4 and F2. TiHo-0906 cells displayed an epithelial/mesenchymal phenotype, and high HMGA2 and CD44 expression. Growth and migration remained comparable during subculturing. Low-passaged cells were two-fold more resistant to doxorubicin than high-passaged cells (IC50: 99.97 nM, and 41.22 nM, respectively). The TiHo-0906 cell line was derived from a poorly differentiated cellular subpopulation of the tumour consistently displaying EMT traits. The cell line presents excellent opportunities for studying EMT on FMCs.

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“…Some of the previously mentioned human and canine PAC and TCC cell lines were characterized in early and some in later passages. However, subculturing-induced gene expression changes were reported to happen within the first ten passages [73], particularly losses of claudin gene expressions [5]. As stable in vitro models are required for repeatable results in further experiments, cell lines were herein characterized in later passages.…”

Section: Plos Onementioning

confidence: 99%

“…Ideally, investigated features of the primary tumor are representative for the tumor type or subtype and stay preserved in the derived cell line [4]. However, clonal selection and adaption to culturing conditions over multiple passages can affect features like gene expressions and sensitivities against chemotherapeutic acting drugs [4][5][6]. Accordingly, the matched characterization of cell lines and respective tissues of origin allows a comprehensive evaluation in which terms a cell line actually represents the tumor entity and can therefore be used as suitable model.…”

Section: Introductionmentioning

confidence: 99%

Characterization of six canine prostate adenocarcinoma and three transitional cell carcinoma cell lines derived from primary tumor tissues as well as metastasis

et al. 2020

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Canine prostate adenocarcinoma (PAC) and transitional cell carcinoma (TCC) of prostate and urinary bladder are highly invasive and metastatic tumors of closely neighbored organs. Cell lines are valuable tools to investigate tumor mechanisms and therapeutic approaches in vitro. PAC in dogs is infrequent, difficult to differentiate from TCC and usually characterized by poor prognosis, enhancing the value of the few available cell lines. However, as cell lines adapt to culturing conditions, a thorough characterization, ideally compared to original tissue, is indispensable. Herein, six canine PAC cell lines and three TCC cell lines were profiled by immunophenotype in comparison to respective original tumor tissues. Three of the six PAC cell lines were derived from primary tumor and metastases of the same patient. Further, two of the three TCC cell lines were derived from TCCs invading into or originating from the prostate. Cell biologic parameters as doubling times and chemoresistances to commonly used drugs in cancer treatment (doxorubicin, carboplatin and meloxicam) were assessed. All cell lines were immunohistochemically close to the respective original tissue. Compared to primary tumor cell lines, metastasis-derived cell lines were more chemoresistant to doxorubicin, but equally susceptive to carboplatin treatment. Two cell lines were multiresistant. COX-2 enzyme activity was demonstrated in all cell lines. However, meloxicam inhibited prostaglandin E2 production in only seven of nine cell lines and did neither influence metabolic activity, nor proliferation. The characterized nine cell lines represent excellent tools to investigate PAC as well as TCC in prostate and urinary bladder of the dog. Furthermore, the profiled paired cell lines from PAC primary tumor and metastasis provide the unique opportunity to investigate metastasis-associated changes PAC cells undergo in tumor progression. The combination of nine differently chemoresistant PAC and TCC cell lines resembles the heterogeneity of canine lower urinary tract cancer. Funding: The authors wish to thank the Studienstiftung des Deutschen Volkes for supporting EMP with a scholarship. This publication was supported by Deutsche Forschungsgemeinschaft and University of PLOS ONE Canine prostate and bladder cancer cell lines derived from primary tumor and metastasis PLOS ONE | https://doi.org/10.Original tissue (P = Prostate, B = urinary bladder, Ln = lymph node); cell lines' names are explained as institution (Tiho = University of Veterinary Medicine Hannover); species (D = dog); tissue origin (Pro = prostate; Urt = urinary tract (urinary bladder)); diagnosis (Adcarc = adenocarcinoma; Carc = carcinoma; Metadcarc = metastasis of an adenocarcinoma); abbreviations of cell lines written in bold; none = cell lines have not been published yet; n.a. = tissue of patient no. 5 is missing, as the patient owners declined surgery and necropsy; � diagnosis by cytology of cells obtained by fine needle aspiration biopsy.

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Longitudinal Claudin Gene Expression Analyses in Canine Mammary Tissues and Thereof Derived Primary Cultures and Cell Lines

Cited by 7 publications

References 50 publications

Analysis of Copy-Number Variations and Feline Mammary Carcinoma Survival

Analysis of Copy-Number Variations and Feline Mammary Carcinoma Survival

TiHo-0906: a new feline mammary cancer cell line with molecular, morphological, and immunocytological characteristics of epithelial to mesenchymal transition

Characterization of six canine prostate adenocarcinoma and three transitional cell carcinoma cell lines derived from primary tumor tissues as well as metastasis

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