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Establishment,functional and genetic characterization of a colon derived large cell neuroendocrine carcinoma cell line

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World Journal of Gastroenterology 2018年第33期24卷 3749-3759页

作者： Michael Gock Christina S Mullins Christine Harnack f.iedrich prall Robert Ramer Anja Goder Oliver H Kramer Ernst Klar Michael Linnebacher Department of General Surgery University of RostockRostock 18055Germany Department of General Surgery Section of Molecular Oncology and ImmunotherapyUniversity of RostockRostock 18055Germany Institute of Pathology University of RostockRostock 18055Germany Institute of Pharmacology University of RostockRostock 18055Germany Institute of Toxicology University Medical Center MainzMainz 55131Germany

AIM To establish cell line and patient-derived xenograft(PDX) models for neuroendocrine carcinomas(NEC) which is highly desirable for gaining insight into tumor development as well as preclinical research includingbiomarker testing and drug response *** Cell line establishment was conducted from direct in vitro culturing of colonic NEC tissue(HROC57). A PDX could also successfully be established from vitally frozen tumor samples. Morphological features, invasive and migratory behavior of the HROC57 cells as well as expression of neuroendocrine markers were vastly analyzed. Phenotypic analysis was done by microscopy and multicolor flow cytometry. The extensive molecular-pathological profiling included mutation analysis, assessment of chromosomal and microsatellite instability; and in addition, fingerprinting(i.e., STR analysis) was performed from the cell line in direct comparison to primary patient-derived tissues and the PDX model established. Drug responsiveness was examined for a panel of chemotherapeutics in clinical use for the treatment of solid *** The established cell line HROC57 showed distinct morphological and molecular features of a poorly differentiated large-cell NEC with KI-67 > 50%. Molecular-pathological analysis revealed a Cp G island promoter methylation positive cell line with microsatellite instability being absent. The following mutation profile was observed: KRAS(wt), BRAf(mut). A high sensitivity to etoposide, cisplatin and 5-fU could be demonstrated while it was more resistant towards rapamycin. CONCLUSION We successfully established and characterized a novel patient-derived NEC cell line in parallel to a PDX model as a useful tool for further analysis of the biological characteristics and for development of novel diagnostic and therapeutic options for NEC.

关键词： Patient-derived tumor model Large cell neuroendocrine carcinoma Individualized medicine

Establishment,functional and genetic characterization of three novel patient-derived rectal cancer cell lines

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World Journal of Gastroenterology 2018年第43期24卷 4880-4892页

作者： Michael Gock Christina S Mullins Carina Bergner f.iedrich prall Robert Ramer Anja Goder Oliver H Kramer falko Lange Bernd J Krause Ernst Klar Michael Linnebacher Department of General Surgery University Medical CenterRostock 18055Germany Section of Molecular Oncology and Immunotherapy University MedicalCenterRostock 18055Germany Department of NuclearMedicine University Medical CenterRostock 18055Germany Institute of Pathology University MedicalCenterRostock 18055Germany Institute of Pharmacology University Medical CenterRostock 18055Germany Institute of Toxicology University Medical Center MainzMainz 55131Germany Oscar-Langendorff-Institute of Physiology University Medical CenterRostock 18055Germany

AIM To establish patient-individual tumor models of rectal cancer for analyses of novel biomarkers, individual response prediction and individual therapy *** Establishment of cell lines was conducted by direct in vitro culturing and in vivo xenografting with subsequent in vitro culturing. Cell lines were in-depth characterized concerning morphological features, invasive and migratory behavior, phenotype, molecular profile including mutational analysis, protein expression, and confirmation of origin by DNA fingerprint. Assessment of chemosensitivity towards an extensive range of current chemotherapeutic drugs and of radiosensitivity was performed including analysis of a combined radioand chemotherapeutic treatment. In addition, glucose metabolism was assessed with 18 f-fluorodeoxyglucose(fDG) and proliferation with 18 *** We describe the establishment of ultra-low passage rectal cancer cell lines of three patients suffering from rectal cancer. Two cell lines(HROC126, HROC284 Met) were established directly from tumor specimens while HROC239 T0 M1 was established subsequent to xenografting of the tumor. Molecular analysis classified all three cell lines as CIMP-0/non-MSI-H(sporadic standard) type. Mutational analysis revealed following mutational profiles: HROC126: APC^(wt), TP53^(wt), KRAS^(wt), BRAf^(wt), PTEN^(wt); HROC239 T0 M1: APC^(mut), P53^(wt), KRAS^(mut), BRAf^(wt), PTEN^(mut) and HROC284 Met: APC^(wt), P53^(mut), KRAS^(mut), BRAf^(wt), PTEN^(mut). All cell lines could be characterized as epithelial(EpCAM+) tumor cells with equivalent morphologic features and comparable growth kinetics. The cell lines displayed a heterogeneous response toward chemotherapy, radiotherapy and their combined application. HROC126 showed a highly radio-resistant phenotype and HROC284 Met was more susceptible to a combined radiochemotherapy than HROC126 and HROC239 T0 M1. Analysis of 18 f-fDG uptake displayed a markedly reduced fDG uptake of all th

关键词： Patient-derived tumor model Rectal cancer ^(18)f-fluorodeoxyglucose ^(18)f-fluorothymidine fOLfOX fOLfIRI Personalized medicine

Establishment and characterization of cell lines from chromosomal instable colorectal cancer

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World Journal of Gastroenterology 2015年第1期21卷 164-176页

作者： Claudia Maletzki Michael Gock Martin Randow Ernst Klar Maja Huehns f.iedrich prall Michael Linnebacher Section of Molecular Oncology and Immunotherapy University of RostockD-18057 RostockGermany Department of General Surgery University of RostockD-18057 RostockGermany Institute of Pathology University of RostockD-18057 RostockGermany

AIM: To generate novel tumor models for preclinicalvalidation of biomarkers that allow drug response prediction and individual therapeutic ***: Cell line establishment was conducted by both direct in vitro culturing and in vivo xenografting followed by in vitro culturing procedure.A comprehensive characterization was subsequently *** included quality control,consisting of the confirmation of human and colorectal cancer(CRC) origin by DNA fingerprint and epithelial cell adhesion molecule(EpC AM) staining,as well as mycoplasma and human virus *** analysis was done by light microscopy and multicolor flow *** examination(β-catenin and cytokeratin staining) was conducted in direct comparison to parental tumor *** molecular-pathological profiling included mutation analysis for CRC-associated driver mutations,assessment of chromosomal and microsatellite instability,and the grade of CpG island ***,an arraybased comparative genomic hybridization analysis was *** responsiveness was assessed for a panel of classical and novel substances in clinical use for the treatment of solid ***,tumorigenicity of the cell lines was tested by xenografting into immunocompromised nude ***: Herein we describe the establishment of three ultra-low passage cell lines from two individual patients suffering from sporadic *** cell line was derived directly from an early stage case(HROC18),whereas two cell lines could be established both direct from patient material and after xenografting from a late stage tumor(HROC32).All cell lines were free of contaminating mycoplasma and *** analysis allowed all cell lines to be classified as chromosomal instable(CIN+).They were aneuploid,with CpG island promoter methylation and microsatellite instability being *** following mutational profile was observed both in the cell lines and the parental t

关键词： Patient-derived tumor model Colorectal cancer Indi

Assessment of Diagnostic Accuracy of Bronchoalveolar Lavage Cytology in the Diagnosis of Lung Tumors and Contribution to the Classification of Non-Small Cell Lung Cancer Entities:A Retrospective Clinocopathological Study

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Open Journal of Pathology 2013年第3期3卷 107-112页

作者： Annette Zimpfer Antonia Polak Andrea Bier Guenther Kundt Jana Kolbel f.iedrich prall Johann C.Virchow Andreas Erbersdobler Institute of Pathology University of RostockRostockGermany Department of Respiratory Medicine University of RostockRostockGermany Institute for Biostatistics and Informatics in Medicine and Ageing Research University of RostockRostockGermany

Background: Due to new therapeutic options in thoracic oncology, the pathological diagnosis of bronchial carcinoma has become more challenging. The majority of bronchial cancer is diagnosed from small biopsy specimens and the diagnosis often based on cytological methods. Aims: In this study, we reevaluated cytologic specimens in order to determine the diagnostic reliability of pulmonary cytopathologic techniques performed in our department. Material and methods: In our center bronchial lavage/bronchoalveolar lavage (BL/BAL) specimens are obtained both before and after forceps biopsy (fB) and subsequently processed. Retrospective data from a period of 60 months were retrieved from the institutional files. Sensitivity, specificity, as well as accuracy of cytological tumor typing were determined using histopathology of fB as gold standard. Also, the diagnostic yield of BL/BAL before and after fB was determined. Results: 678 cases were retrieved from the institutional files. The sensitivity and specificity of cytology were 83.0% and 83.4%, respectively. By fB in 3.9% of cytologically diagnosed non-small cell lung carcinomas (NSCLC) a histological assignment to a NSCLC entity was not possible. Conclusions: Cytology is a reliable diagnostic tool in the diagnosis of lung malignancies. High diagnostic accuracy is achieved by a combination of BL/BAL before and after fB. The diagnostic yield of BL/BAL after fB was significantly higher than BL/BAL before fB. Subsequent tumor typing of cytologically diagnosed NSCLC was feasible in more than 95% of cases.

关键词： Lavage Cytology Accuracy Lung Cancer NSCLC Classification

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适应现代价值创造重塑报告准则

中国资产评估 2021年第12期 68-72页

作者：乔元芳(编译) 凯文·普拉尔上海市财政局上海200030 国际评估准则理事会

随着知识经济的到来,经济驱动因素和商业模式发生了根本性变化,无形资产在经济和企业成长与发展中的作用日益凸显。但是,财务报表中未确认的无形资产占比越来越大,公司市值与净资产的差距越来越明显,投资者越来越倾向于通过其他渠道获... 详细信息

随着知识经济的到来,经济驱动因素和商业模式发生了根本性变化,无形资产在经济和企业成长与发展中的作用日益凸显。但是,财务报表中未确认的无形资产占比越来越大,公司市值与净资产的差距越来越明显,投资者越来越倾向于通过其他渠道获得决策所需要的信息,财务报表的相关性逐步降低,研究探索新型的无形资产会计框架迫在眉睫。

关键词：无形资产确认披露