miR-21 expression analysis in budding colon cancer cells by confocal slide scanning microscopy

Kirsten Nguyen Knudsen*, Jan Lindebjerg, Alexandra Kalmár, Béla Molnár, Flemming Brandt Sørensen, Torben Frøstrup Hansen, Boye Schnack Nielsen

*Kontaktforfatter for dette arbejde

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Resumé

MicroRNA-21 (miR-21) expression in stromal fibroblastic cells in colorectal cancer is well-documented, whereas miR-21 expression in tumor budding cells (TBCs) is poorly described. TBCs are locally invasive carcinoma cells with increased metastatic properties and characteristics of epithelial to mesenchymal transition. This study was conducted to better characterize the expression of miR-21 in TBCs. First, chromogenic miR-21 in situ hybridization (ISH) staining was performed in 58 colon adenocarcinomas with evident TBCs. Then, to obtain unambiguous identification of miR-21 in the TBCs, twenty cases were selected for an additional multiplex fluorescence analysis combining miR-21 ISH with cytokeratin and laminin-5γ2 immunofluorescence. Employing confocal slide scanning microscopy, comprehensive digital images of the invasive front (10–40 mm2) were obtained from 16 of the 20 cases, and miR-21 expression was evaluated in cytokeratin-positive TBCs. The high resolution of the confocal digital slide images allowed a detailed examination of the confocal stacks of the multiplex-stained tissue sections. The cases with the highest fraction of miR-21 positive TBCs were all stage III cancers defined by the presence of regional lymph node metastasis. Some of the miR-21 positive TBCs were also laminin-5γ2 positive. The confocal image stacks also revealed that some TBCs were actually directly connected to malignant glands. In conclusion, miR-21 expression was unambiguously identified in TBCs by evaluation of digital slides obtained by confocal slide scanning microscopy. In addition, the digital confocal slides provided a more detailed understanding of local cancer cell invasion by allowing evaluation of the cell structures in three dimensions.

OriginalsprogEngelsk
TidsskriftClinical and Experimental Metastasis
Vol/bind35
Udgave nummer8
Sider (fra-til)819–830
ISSN0262-0898
DOI
StatusUdgivet - 2018

Fingeraftryk

MicroRNAs
Colonic Neoplasms
Microscopy
Neoplasms
Laminin
Stromal Cells
Colorectal Neoplasms
Colon
Fluorescence
Lymph Nodes

Citer dette

Knudsen, Kirsten Nguyen ; Lindebjerg, Jan ; Kalmár, Alexandra ; Molnár, Béla ; Sørensen, Flemming Brandt ; Hansen, Torben Frøstrup ; Nielsen, Boye Schnack. / miR-21 expression analysis in budding colon cancer cells by confocal slide scanning microscopy. I: Clinical and Experimental Metastasis. 2018 ; Bind 35, Nr. 8. s. 819–830.
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title = "miR-21 expression analysis in budding colon cancer cells by confocal slide scanning microscopy",
abstract = "MicroRNA-21 (miR-21) expression in stromal fibroblastic cells in colorectal cancer is well-documented, whereas miR-21 expression in tumor budding cells (TBCs) is poorly described. TBCs are locally invasive carcinoma cells with increased metastatic properties and characteristics of epithelial to mesenchymal transition. This study was conducted to better characterize the expression of miR-21 in TBCs. First, chromogenic miR-21 in situ hybridization (ISH) staining was performed in 58 colon adenocarcinomas with evident TBCs. Then, to obtain unambiguous identification of miR-21 in the TBCs, twenty cases were selected for an additional multiplex fluorescence analysis combining miR-21 ISH with cytokeratin and laminin-5γ2 immunofluorescence. Employing confocal slide scanning microscopy, comprehensive digital images of the invasive front (10–40 mm2) were obtained from 16 of the 20 cases, and miR-21 expression was evaluated in cytokeratin-positive TBCs. The high resolution of the confocal digital slide images allowed a detailed examination of the confocal stacks of the multiplex-stained tissue sections. The cases with the highest fraction of miR-21 positive TBCs were all stage III cancers defined by the presence of regional lymph node metastasis. Some of the miR-21 positive TBCs were also laminin-5γ2 positive. The confocal image stacks also revealed that some TBCs were actually directly connected to malignant glands. In conclusion, miR-21 expression was unambiguously identified in TBCs by evaluation of digital slides obtained by confocal slide scanning microscopy. In addition, the digital confocal slides provided a more detailed understanding of local cancer cell invasion by allowing evaluation of the cell structures in three dimensions.",
keywords = "Colon cancer, Confocal slide scanning microscopy, Digital imaging, MicroRNA-21, Multiplex fluorescence, Tumor budding cells",
author = "Knudsen, {Kirsten Nguyen} and Jan Lindebjerg and Alexandra Kalm{\'a}r and B{\'e}la Moln{\'a}r and S{\o}rensen, {Flemming Brandt} and Hansen, {Torben Fr{\o}strup} and Nielsen, {Boye Schnack}",
year = "2018",
doi = "10.1007/s10585-018-9945-3",
language = "English",
volume = "35",
pages = "819–830",
journal = "Clinical and Experimental Metastasis",
issn = "0262-0898",
publisher = "Springer",
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miR-21 expression analysis in budding colon cancer cells by confocal slide scanning microscopy. / Knudsen, Kirsten Nguyen; Lindebjerg, Jan; Kalmár, Alexandra; Molnár, Béla; Sørensen, Flemming Brandt; Hansen, Torben Frøstrup; Nielsen, Boye Schnack.

I: Clinical and Experimental Metastasis, Bind 35, Nr. 8, 2018, s. 819–830.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - miR-21 expression analysis in budding colon cancer cells by confocal slide scanning microscopy

AU - Knudsen, Kirsten Nguyen

AU - Lindebjerg, Jan

AU - Kalmár, Alexandra

AU - Molnár, Béla

AU - Sørensen, Flemming Brandt

AU - Hansen, Torben Frøstrup

AU - Nielsen, Boye Schnack

PY - 2018

Y1 - 2018

N2 - MicroRNA-21 (miR-21) expression in stromal fibroblastic cells in colorectal cancer is well-documented, whereas miR-21 expression in tumor budding cells (TBCs) is poorly described. TBCs are locally invasive carcinoma cells with increased metastatic properties and characteristics of epithelial to mesenchymal transition. This study was conducted to better characterize the expression of miR-21 in TBCs. First, chromogenic miR-21 in situ hybridization (ISH) staining was performed in 58 colon adenocarcinomas with evident TBCs. Then, to obtain unambiguous identification of miR-21 in the TBCs, twenty cases were selected for an additional multiplex fluorescence analysis combining miR-21 ISH with cytokeratin and laminin-5γ2 immunofluorescence. Employing confocal slide scanning microscopy, comprehensive digital images of the invasive front (10–40 mm2) were obtained from 16 of the 20 cases, and miR-21 expression was evaluated in cytokeratin-positive TBCs. The high resolution of the confocal digital slide images allowed a detailed examination of the confocal stacks of the multiplex-stained tissue sections. The cases with the highest fraction of miR-21 positive TBCs were all stage III cancers defined by the presence of regional lymph node metastasis. Some of the miR-21 positive TBCs were also laminin-5γ2 positive. The confocal image stacks also revealed that some TBCs were actually directly connected to malignant glands. In conclusion, miR-21 expression was unambiguously identified in TBCs by evaluation of digital slides obtained by confocal slide scanning microscopy. In addition, the digital confocal slides provided a more detailed understanding of local cancer cell invasion by allowing evaluation of the cell structures in three dimensions.

AB - MicroRNA-21 (miR-21) expression in stromal fibroblastic cells in colorectal cancer is well-documented, whereas miR-21 expression in tumor budding cells (TBCs) is poorly described. TBCs are locally invasive carcinoma cells with increased metastatic properties and characteristics of epithelial to mesenchymal transition. This study was conducted to better characterize the expression of miR-21 in TBCs. First, chromogenic miR-21 in situ hybridization (ISH) staining was performed in 58 colon adenocarcinomas with evident TBCs. Then, to obtain unambiguous identification of miR-21 in the TBCs, twenty cases were selected for an additional multiplex fluorescence analysis combining miR-21 ISH with cytokeratin and laminin-5γ2 immunofluorescence. Employing confocal slide scanning microscopy, comprehensive digital images of the invasive front (10–40 mm2) were obtained from 16 of the 20 cases, and miR-21 expression was evaluated in cytokeratin-positive TBCs. The high resolution of the confocal digital slide images allowed a detailed examination of the confocal stacks of the multiplex-stained tissue sections. The cases with the highest fraction of miR-21 positive TBCs were all stage III cancers defined by the presence of regional lymph node metastasis. Some of the miR-21 positive TBCs were also laminin-5γ2 positive. The confocal image stacks also revealed that some TBCs were actually directly connected to malignant glands. In conclusion, miR-21 expression was unambiguously identified in TBCs by evaluation of digital slides obtained by confocal slide scanning microscopy. In addition, the digital confocal slides provided a more detailed understanding of local cancer cell invasion by allowing evaluation of the cell structures in three dimensions.

KW - Colon cancer

KW - Confocal slide scanning microscopy

KW - Digital imaging

KW - MicroRNA-21

KW - Multiplex fluorescence

KW - Tumor budding cells

U2 - 10.1007/s10585-018-9945-3

DO - 10.1007/s10585-018-9945-3

M3 - Journal article

VL - 35

SP - 819

EP - 830

JO - Clinical and Experimental Metastasis

JF - Clinical and Experimental Metastasis

SN - 0262-0898

IS - 8

ER -