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Expression profile of microenvironment of morphological structures in breast cancer

ABSTRACT: The aim of this work is to compare the expression profiles of the microenvironment of various morphological structures of luminal breast cancer obtained by laser microdissection. For this, sections of primary breast carcinoma were stained according to the RNA-preserving protocol, fragments of the stroma around alveolar, trabecular, solid structures and single tumor cells were isolated using a laser capture microdissection, and then RNA-sequencing was performed using Illumina NextSeq500. Our study presents the first analysis of DEGs and activated signaling pathways of the microenvironment of various morphological structures of breast cancer.

ORGANISM(S): Homo sapiens

PROVIDER: GSE184196 | GEO | 2021/09/22

REPOSITORIES: GEO

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Project description:The classical sacrococcygeal chordoma tumor presents with a typical morphology of lobulated myxoid tumor tissue with cords, strands and nests of tumor cells consisting of small non-vacuolated cells, intermediate cells with a wide range of vacuolization and large heavily vacuolated (physaliferous) cells. Because of its rare incidence, lack of suited model systems and technical limitations analysis was only performed on bulk tumor mass neglecting its heterogeneous composition. We aimed at elucidating the differences between small non-vacuolated and large physaliferous cells on the genomic and transcriptomic level. Secondly, we intended to clarify whether the observed cell types are derived from genetically distinct clones or rather represent different phenotypes. Using the chordoma cell line MUG-Chor1 we monitored morphological changes via time lapse experiments. We isolated pure fractions of each phenotype by means of laser microdissection or micromanipulation allowing phenotype-specific analysis. Pools of 100 cells each were genetically profiled after whole genome amplification by array comparative genomic hybridization. For expression analysis 20 cells each were subjected to whole transcriptom amplification, forwarded to RNA microarray analysis and qRT-PCR. Time lapse analysis unveiled small non-vacuolated cells to develop into large physaliferous cells via intermediate cells containing an increasing amount of vacuoles. Furthermore, we showed small and large physaliferous cells to proliferate at the same rate but intermediate cells to be the most proliferating cell phenotype. Small non-vacuolated and large physaliferous cells showed identical copy number variations. Despite their obvious morphological disparities we detected only modest changes in over all gene expression. However, verification of candidate genes yielded significant up-regulation of ALG11 (700-fold), PPP2CB (18.6-fold), and UCHL3 (18.7-fold) in large physaliferous cells. Of two different cell types (large and small MUG-Chor1 cells) we analysed each in triplicates. In total 6 cell pools were analysed.

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Expression profile of microenvironment of morphological structures in breast cancer

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