Mesenchymal change and drug resistance in neuroblastoma

Jessica A. Naiditch, Chunfa Jie, Timothy B. Lautz, Songtao Yu, Sandra Clark, Dimitry Voronov, Fei Chu, Mary Beth Madonna

Research output: Contribution to journalArticle

Abstract

Background: Metastatic initiation has many phenotypic similarities to epithelial-tomesenchymal transition, including loss of cell-cell adhesion, increased invasiveness, and increased cell mobility. We have previously demonstrated that drug resistance is associated with a metastatic phenotype in neuroblastoma (NB). The purpose of this project was to determine if the development of doxorubicin resistance is associated with characteristics of mesenchymal change in human NB cells. Materials and methods: Total RNA was isolated from wild type (WT) and doxorubicinresistant (DoxR) human NB cell lines (SK-N-SH and SK-N-BE(2)C) and analyzed using the Illumina Human HT-12 version 4 Expression BeadChip. Differentially expressed genes (DEGs) were identified. Volcano plots and heat maps were generated. Genes of interest with a fold change in expression >1.5 and an adjusted P <0.1 were analyzed. Immunofluorescence (IF) and Western blot analysis confirmed microarray results of interest. Matrigel invasion assay and migration wounding assays were performed. Results: Volcano plots and heat maps visually demonstrated a similar pattern of DEGs in the SK-N-SH and SK-N-BE(2)C DoxR cell lines relative to their parental WT lines. Venn diagramming revealed 1594 DEGs common to both DoxR cell lines relative to their parental cell lines. Network analysis pointed to several significantly upregulated epithelial-tomesenchymal transition pathways, through TGF-beta pathways via RhoA, PI3K, and ILK and via SMADs, as well as via notch signaling pathways. DoxR cell lines displayed a more invasive phenotype than respective WT cell lines. Conclusions: Human SK-N-SH and SK-N-BE(2)C NB cells display characteristics of mesenchymal change via multiple pathways in the transition to a drug-resistant state.

Original languageEnglish (US)
Pages (from-to)279-288
Number of pages10
JournalJournal of Surgical Research
Volume193
Issue number1
DOIs
StatePublished - 2015
Externally publishedYes

Fingerprint

Neuroblastoma
Drug Resistance
Cell Line
Genes
Hot Temperature
Phenotype
Microarray Analysis
Phosphatidylinositol 3-Kinases
Cell Adhesion
Transforming Growth Factor beta
Doxorubicin
Fluorescent Antibody Technique
Western Blotting
RNA
Pharmaceutical Preparations

Keywords

  • Drug resistance
  • Long-term drug exposure
  • Mesenchymal change
  • Neuroblastoma

ASJC Scopus subject areas

  • Surgery
  • Medicine(all)

Cite this

Naiditch, J. A., Jie, C., Lautz, T. B., Yu, S., Clark, S., Voronov, D., ... Madonna, M. B. (2015). Mesenchymal change and drug resistance in neuroblastoma. Journal of Surgical Research, 193(1), 279-288. https://doi.org/10.1016/j.jss.2014.07.018

Mesenchymal change and drug resistance in neuroblastoma. / Naiditch, Jessica A.; Jie, Chunfa; Lautz, Timothy B.; Yu, Songtao; Clark, Sandra; Voronov, Dimitry; Chu, Fei; Madonna, Mary Beth.

In: Journal of Surgical Research, Vol. 193, No. 1, 2015, p. 279-288.

Research output: Contribution to journalArticle

Naiditch, JA, Jie, C, Lautz, TB, Yu, S, Clark, S, Voronov, D, Chu, F & Madonna, MB 2015, 'Mesenchymal change and drug resistance in neuroblastoma', Journal of Surgical Research, vol. 193, no. 1, pp. 279-288. https://doi.org/10.1016/j.jss.2014.07.018
Naiditch JA, Jie C, Lautz TB, Yu S, Clark S, Voronov D et al. Mesenchymal change and drug resistance in neuroblastoma. Journal of Surgical Research. 2015;193(1):279-288. https://doi.org/10.1016/j.jss.2014.07.018
Naiditch, Jessica A. ; Jie, Chunfa ; Lautz, Timothy B. ; Yu, Songtao ; Clark, Sandra ; Voronov, Dimitry ; Chu, Fei ; Madonna, Mary Beth. / Mesenchymal change and drug resistance in neuroblastoma. In: Journal of Surgical Research. 2015 ; Vol. 193, No. 1. pp. 279-288.
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abstract = "Background: Metastatic initiation has many phenotypic similarities to epithelial-tomesenchymal transition, including loss of cell-cell adhesion, increased invasiveness, and increased cell mobility. We have previously demonstrated that drug resistance is associated with a metastatic phenotype in neuroblastoma (NB). The purpose of this project was to determine if the development of doxorubicin resistance is associated with characteristics of mesenchymal change in human NB cells. Materials and methods: Total RNA was isolated from wild type (WT) and doxorubicinresistant (DoxR) human NB cell lines (SK-N-SH and SK-N-BE(2)C) and analyzed using the Illumina Human HT-12 version 4 Expression BeadChip. Differentially expressed genes (DEGs) were identified. Volcano plots and heat maps were generated. Genes of interest with a fold change in expression >1.5 and an adjusted P <0.1 were analyzed. Immunofluorescence (IF) and Western blot analysis confirmed microarray results of interest. Matrigel invasion assay and migration wounding assays were performed. Results: Volcano plots and heat maps visually demonstrated a similar pattern of DEGs in the SK-N-SH and SK-N-BE(2)C DoxR cell lines relative to their parental WT lines. Venn diagramming revealed 1594 DEGs common to both DoxR cell lines relative to their parental cell lines. Network analysis pointed to several significantly upregulated epithelial-tomesenchymal transition pathways, through TGF-beta pathways via RhoA, PI3K, and ILK and via SMADs, as well as via notch signaling pathways. DoxR cell lines displayed a more invasive phenotype than respective WT cell lines. Conclusions: Human SK-N-SH and SK-N-BE(2)C NB cells display characteristics of mesenchymal change via multiple pathways in the transition to a drug-resistant state.",
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AU - Voronov, Dimitry

AU - Chu, Fei

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N2 - Background: Metastatic initiation has many phenotypic similarities to epithelial-tomesenchymal transition, including loss of cell-cell adhesion, increased invasiveness, and increased cell mobility. We have previously demonstrated that drug resistance is associated with a metastatic phenotype in neuroblastoma (NB). The purpose of this project was to determine if the development of doxorubicin resistance is associated with characteristics of mesenchymal change in human NB cells. Materials and methods: Total RNA was isolated from wild type (WT) and doxorubicinresistant (DoxR) human NB cell lines (SK-N-SH and SK-N-BE(2)C) and analyzed using the Illumina Human HT-12 version 4 Expression BeadChip. Differentially expressed genes (DEGs) were identified. Volcano plots and heat maps were generated. Genes of interest with a fold change in expression >1.5 and an adjusted P <0.1 were analyzed. Immunofluorescence (IF) and Western blot analysis confirmed microarray results of interest. Matrigel invasion assay and migration wounding assays were performed. Results: Volcano plots and heat maps visually demonstrated a similar pattern of DEGs in the SK-N-SH and SK-N-BE(2)C DoxR cell lines relative to their parental WT lines. Venn diagramming revealed 1594 DEGs common to both DoxR cell lines relative to their parental cell lines. Network analysis pointed to several significantly upregulated epithelial-tomesenchymal transition pathways, through TGF-beta pathways via RhoA, PI3K, and ILK and via SMADs, as well as via notch signaling pathways. DoxR cell lines displayed a more invasive phenotype than respective WT cell lines. Conclusions: Human SK-N-SH and SK-N-BE(2)C NB cells display characteristics of mesenchymal change via multiple pathways in the transition to a drug-resistant state.

AB - Background: Metastatic initiation has many phenotypic similarities to epithelial-tomesenchymal transition, including loss of cell-cell adhesion, increased invasiveness, and increased cell mobility. We have previously demonstrated that drug resistance is associated with a metastatic phenotype in neuroblastoma (NB). The purpose of this project was to determine if the development of doxorubicin resistance is associated with characteristics of mesenchymal change in human NB cells. Materials and methods: Total RNA was isolated from wild type (WT) and doxorubicinresistant (DoxR) human NB cell lines (SK-N-SH and SK-N-BE(2)C) and analyzed using the Illumina Human HT-12 version 4 Expression BeadChip. Differentially expressed genes (DEGs) were identified. Volcano plots and heat maps were generated. Genes of interest with a fold change in expression >1.5 and an adjusted P <0.1 were analyzed. Immunofluorescence (IF) and Western blot analysis confirmed microarray results of interest. Matrigel invasion assay and migration wounding assays were performed. Results: Volcano plots and heat maps visually demonstrated a similar pattern of DEGs in the SK-N-SH and SK-N-BE(2)C DoxR cell lines relative to their parental WT lines. Venn diagramming revealed 1594 DEGs common to both DoxR cell lines relative to their parental cell lines. Network analysis pointed to several significantly upregulated epithelial-tomesenchymal transition pathways, through TGF-beta pathways via RhoA, PI3K, and ILK and via SMADs, as well as via notch signaling pathways. DoxR cell lines displayed a more invasive phenotype than respective WT cell lines. Conclusions: Human SK-N-SH and SK-N-BE(2)C NB cells display characteristics of mesenchymal change via multiple pathways in the transition to a drug-resistant state.

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