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

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

Research output: Contribution to journalArticle

4 Citations (Scopus)

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.

Original languageEnglish
JournalClinical and Experimental Metastasis
DOIs
Publication statusAccepted/In press - Jan 1 2018

Fingerprint

MicroRNAs
Colonic Neoplasms
Microscopy
Neoplasms
Laminin
Keratins
In Situ Hybridization
Epithelial-Mesenchymal Transition
Stromal Cells
Fluorescent Antibody Technique
Colorectal Neoplasms
Colon
Adenocarcinoma
Fluorescence
Lymph Nodes
Staining and Labeling
Neoplasm Metastasis
Carcinoma

Keywords

  • Colon cancer
  • Confocal slide scanning microscopy
  • Digital imaging
  • MicroRNA-21
  • Multiplex fluorescence
  • Tumor budding cells

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

miR-21 expression analysis in budding colon cancer cells by confocal slide scanning microscopy. / Knudsen, Kirsten Nguyen; Lindebjerg, Jan; Kalmár, Alexandra; Molnár, B.; Sørensen, Flemming Brandt; Hansen, Torben Frøstrup; Nielsen, Boye Schnack.

In: Clinical and Experimental Metastasis, 01.01.2018.

Research output: Contribution to journalArticle

Knudsen, Kirsten Nguyen ; Lindebjerg, Jan ; Kalmár, Alexandra ; Molnár, B. ; Sørensen, Flemming Brandt ; Hansen, Torben Frøstrup ; Nielsen, Boye Schnack. / miR-21 expression analysis in budding colon cancer cells by confocal slide scanning microscopy. In: Clinical and Experimental Metastasis. 2018.
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