Uniformity under in vitro conditions: Changes in the phenotype
of cancer cell lines derived from different medulloblastoma
subgroups.

J 2017

Uniformity under in vitro conditions: Changes in the phenotype of cancer cell lines derived from different medulloblastoma subgroups.

CHLAPEK, Petr, Karel ZITTERBART, Leoš KŘEN, Lenka FILIPOVÁ, Jaroslav ŠTĚRBA et. al.

Basic information

Original name

Uniformity under in vitro conditions: Changes in the phenotype of cancer cell lines derived from different medulloblastoma subgroups.

Authors

CHLAPEK, Petr (203 Czech Republic, belonging to the institution), Karel ZITTERBART (203 Czech Republic, belonging to the institution), Leoš KŘEN (203 Czech Republic), Lenka FILIPOVÁ (203 Czech Republic, belonging to the institution), Jaroslav ŠTĚRBA (203 Czech Republic, belonging to the institution) and Renata VESELSKÁ (203 Czech Republic, guarantor, belonging to the institution)

Edition

PLOS ONE, SAN FRANCISCO, PUBLIC LIBRARY SCIENCE, 2017, 1932-6203

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10601 Cell biology

Country of publisher

United States of America

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL

Impact factor

Impact factor: 2.766

RIV identification code

RIV/00216224:14310/17:00099806

Organization unit

Faculty of Science

DOI

http://dx.doi.org/10.1371/journal.pone.0172552

UT WoS

000394682400055

Keywords in English

medulloblastoma; molecular subgroups; primary tumors; cell lines; beta-catenin; filamin A; GAB1; YAP1; immunodetection; protein markers

Abstract

V originále

Medulloblastoma comprises four main subgroups (WNT, SHH, Group 3 and Group 4) originally defined by transcriptional profiling. In primary medulloblastoma tissues, these groups are thought to be distinguishable using the immunohistochemical detection of beta-catenin, filamin A, GAB1 and YAP1 protein markers. To investigate the utility of these markers for in vitro studies using medulloblastoma cell lines, immunoblotting and indirect immunofluorescence were employed for the detection of beta-catenin, filamin A, GAB1 and YAP1 in both DAOY and D283 Med reference cell lines and the panel of six medulloblastoma cell lines derived in our laboratory from the primary tumor tissues of known molecular subgroups. Immunohistochemical detection of these markers was performed on formalin-fixed paraffin-embedded tissue of the matching primary tumors. The results revealed substantial divergences between the primary tumor tissues and matching cell lines in the immunoreactivity pattern of medulloblastoma-subgroup-specific protein markers. Regardless of the molecular subgroup of the primary tumor, all six patient-derived medulloblastoma cell lines exhibited a uniform phenotype: immunofluorescence showed the nuclear localization of YAP1, accompanied by strong cytoplasmic positivity for beta-catenin and filamin A, as well as weak positivity for GAB1. The same immunoreactivity pattern was also found in both DAOY and D283 Med reference medulloblastoma cell lines. Therefore, we can conclude that various medulloblastoma cell lines tend to exhibit the same characteristics of protein marker expression under standard in vitro conditions. Such a finding emphasizes the importance of the analyses of primary tumors in clinically oriented medulloblastoma research and the urgent need to develop in vitro models of improved clinical relevance, such as 3D cultures and organotypic slice cultures.

Citovat

CHLAPEK, Petr, Karel ZITTERBART, Leoš KŘEN, Lenka FILIPOVÁ, Jaroslav ŠTĚRBA and Renata VESELSKÁ. Uniformity under in vitro conditions: Changes in the phenotype of cancer cell lines derived from different medulloblastoma subgroups. PLOS ONE. SAN FRANCISCO: PUBLIC LIBRARY SCIENCE, 2017, vol. 12, No 2, p. nestránkováno, 14 pp. ISSN 1932-6203. Available from: https://dx.doi.org/10.1371/journal.pone.0172552.

@article{1406174,
   author = {Chlapek, Petr and Zitterbart, Karel and Křen, Leoš and Filipová, Lenka and Štěrba, Jaroslav and Veselská, Renata},
   article_location = {SAN FRANCISCO},
   article_number = {2},
   doi = {http://dx.doi.org/10.1371/journal.pone.0172552},
   keywords = {medulloblastoma; molecular subgroups; primary tumors; cell lines; beta-catenin; filamin A; GAB1; YAP1; immunodetection; protein markers},
   language = {eng},
   issn = {1932-6203},
   journal = {PLOS ONE},
   title = {Uniformity under in vitro conditions: Changes in the phenotype of cancer cell lines derived from different medulloblastoma subgroups.},
   url = {http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0172552},
   volume = {12},
   year = {2017}
}

TY  - JOUR
ID  - 1406174
AU  - Chlapek, Petr - Zitterbart, Karel - Křen, Leoš - Filipová, Lenka - Štěrba, Jaroslav - Veselská, Renata
PY  - 2017
TI  - Uniformity under in vitro conditions: Changes in the phenotype of cancer cell lines derived from different medulloblastoma subgroups.
JF  - PLOS ONE
VL  - 12
IS  - 2
SP  - nestránkováno
EP  - nestránkováno
PB  - PUBLIC LIBRARY SCIENCE
SN  - 19326203
KW  - medulloblastoma
KW  - molecular subgroups
KW  - primary tumors
KW  - cell lines
KW  - beta-catenin
KW  - filamin A
KW  - GAB1
KW  - YAP1
KW  - immunodetection
KW  - protein markers
UR  - http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0172552
L2  - http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0172552
N2  - Medulloblastoma comprises four main subgroups (WNT, SHH, Group 3 and Group 4) originally defined by transcriptional profiling. In primary medulloblastoma tissues, these groups are thought to be distinguishable using the immunohistochemical detection of beta-catenin, filamin A, GAB1 and YAP1 protein markers. To investigate the utility of these markers for in vitro studies using medulloblastoma cell lines, immunoblotting and indirect immunofluorescence were employed for the detection of beta-catenin, filamin A, GAB1 and YAP1 in both DAOY and D283 Med reference cell lines and the panel of six medulloblastoma cell lines derived in our laboratory from the primary tumor tissues of known molecular subgroups. Immunohistochemical detection of these markers was performed on formalin-fixed paraffin-embedded tissue of the matching primary tumors. The results revealed substantial divergences between the primary tumor tissues and matching cell lines in the immunoreactivity pattern of medulloblastoma-subgroup-specific protein markers. Regardless of the molecular subgroup of the primary tumor, all six patient-derived medulloblastoma cell lines exhibited a uniform phenotype: immunofluorescence showed the nuclear localization of YAP1, accompanied by strong cytoplasmic positivity for beta-catenin and filamin A, as well as weak positivity for GAB1. The same immunoreactivity pattern was also found in both DAOY and D283 Med reference medulloblastoma cell lines. Therefore, we can conclude that various medulloblastoma cell lines tend to exhibit the same characteristics of protein marker expression under standard in vitro conditions. Such a finding emphasizes the importance of the analyses of primary tumors in clinically oriented medulloblastoma research and the urgent need to develop in vitro models of improved clinical relevance, such as 3D cultures and organotypic slice cultures.
ER  -

CHLAPEK, Petr, Karel ZITTERBART, Leoš KŘEN, Lenka FILIPOVÁ, Jaroslav ŠTĚRBA and Renata VESELSKÁ. Uniformity under in vitro conditions: Changes in the phenotype of cancer cell lines derived from different medulloblastoma subgroups. \textit{PLOS ONE}. SAN FRANCISCO: PUBLIC LIBRARY SCIENCE, 2017, vol.~12, No~2, p.~nestránkováno, 14 pp. ISSN~1932-6203. Available from: https://dx.doi.org/10.1371/journal.pone.0172552.

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