Microarray analysis reveals potential target genes of NF-κB2/p52 in LNCaP prostate cancer cells

Nagalakshmi Nadiminty, Smitha Dutt, Clifford G Tepper, Allen C Gao

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

PURPOSE. Our previous studies showed that NF-κB2/p52 is involved in the castration-resistant growth of the androgen-sensitive LNCaP prostate cancer cells. The role of NF-κB2/p52 in lymphomagenesis has been studied extensively, but its target genes in other cancers remain unknown. In order to identify genes potentially regulated by p52 in prostate cancer cells, we performed a genome-wide microarray analysis of genes differentially up- or down-regulated by the overexpression of p52 by adenoviral-mediated gene delivery in LNCaP cells. EXPERIMENTAL DESIGN. Total RNAs from vector control-infected and Adeno-p52-infected LNCaP cells were used to prepare cDNAs, which were hybridized to the Whole Genome Human 44k Microarray chips (Agilent Technologies). Data analysis was performed using GeneSpring and Ingenuity Pathway Analysis software. Validation of microarray results was performed by real-time quantitative RT-PCR and Western blot analyses. RESULTS. Expression of ∼130 genes was differentially upregulated by >5-fold, whereas ∼60 genes were differentially downregulated by >2-fold in p52-expressing LNCaP cells. Pathway analysis revealed that the upregulated genes belong to functional categories like cell growth and proliferation, cellular movement, cell-to-cell signaling and interaction, cancer, cell cycle, etc., whereas the downregulated genes were represented by functional categories like cell movement, antigen presentation, and cell death. Six of the top upregulated genes including annexin A2, PLAU, RND3, Twist2, VEGFC, and CXCL1 were validated by real-time PCR and Western blot analysis. CONCLUSIONS. This study provides a comprehensive analysis of genes potentially regulated by NF-κB2/p52 in the LNCaP prostate cancer cell line and provides a rationale for the induction of castration-resistant growth by p52 in LNCaP cells.

Original languageEnglish (US)
Pages (from-to)276-287
Number of pages12
JournalProstate
Volume70
Issue number3
DOIs
StatePublished - Feb 15 2010

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Microarray Analysis
Prostatic Neoplasms
Genes
Castration
Real-Time Polymerase Chain Reaction
Down-Regulation
Growth
Western Blotting
Annexin A2
Antigen Presentation
Human Genome
Cell Communication
Androgens
Cell Movement
Neoplasms
Cell Cycle
Cell Death
Software
Complementary DNA
Cell Proliferation

Keywords

  • LNCaP
  • Microarray
  • NF-κB2/p52
  • Prostate

ASJC Scopus subject areas

  • Urology
  • Oncology

Cite this

Microarray analysis reveals potential target genes of NF-κB2/p52 in LNCaP prostate cancer cells. / Nadiminty, Nagalakshmi; Dutt, Smitha; Tepper, Clifford G; Gao, Allen C.

In: Prostate, Vol. 70, No. 3, 15.02.2010, p. 276-287.

Research output: Contribution to journalArticle

Nadiminty, Nagalakshmi ; Dutt, Smitha ; Tepper, Clifford G ; Gao, Allen C. / Microarray analysis reveals potential target genes of NF-κB2/p52 in LNCaP prostate cancer cells. In: Prostate. 2010 ; Vol. 70, No. 3. pp. 276-287.
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abstract = "PURPOSE. Our previous studies showed that NF-κB2/p52 is involved in the castration-resistant growth of the androgen-sensitive LNCaP prostate cancer cells. The role of NF-κB2/p52 in lymphomagenesis has been studied extensively, but its target genes in other cancers remain unknown. In order to identify genes potentially regulated by p52 in prostate cancer cells, we performed a genome-wide microarray analysis of genes differentially up- or down-regulated by the overexpression of p52 by adenoviral-mediated gene delivery in LNCaP cells. EXPERIMENTAL DESIGN. Total RNAs from vector control-infected and Adeno-p52-infected LNCaP cells were used to prepare cDNAs, which were hybridized to the Whole Genome Human 44k Microarray chips (Agilent Technologies). Data analysis was performed using GeneSpring and Ingenuity Pathway Analysis software. Validation of microarray results was performed by real-time quantitative RT-PCR and Western blot analyses. RESULTS. Expression of ∼130 genes was differentially upregulated by >5-fold, whereas ∼60 genes were differentially downregulated by >2-fold in p52-expressing LNCaP cells. Pathway analysis revealed that the upregulated genes belong to functional categories like cell growth and proliferation, cellular movement, cell-to-cell signaling and interaction, cancer, cell cycle, etc., whereas the downregulated genes were represented by functional categories like cell movement, antigen presentation, and cell death. Six of the top upregulated genes including annexin A2, PLAU, RND3, Twist2, VEGFC, and CXCL1 were validated by real-time PCR and Western blot analysis. CONCLUSIONS. This study provides a comprehensive analysis of genes potentially regulated by NF-κB2/p52 in the LNCaP prostate cancer cell line and provides a rationale for the induction of castration-resistant growth by p52 in LNCaP cells.",
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AU - Gao, Allen C

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N2 - PURPOSE. Our previous studies showed that NF-κB2/p52 is involved in the castration-resistant growth of the androgen-sensitive LNCaP prostate cancer cells. The role of NF-κB2/p52 in lymphomagenesis has been studied extensively, but its target genes in other cancers remain unknown. In order to identify genes potentially regulated by p52 in prostate cancer cells, we performed a genome-wide microarray analysis of genes differentially up- or down-regulated by the overexpression of p52 by adenoviral-mediated gene delivery in LNCaP cells. EXPERIMENTAL DESIGN. Total RNAs from vector control-infected and Adeno-p52-infected LNCaP cells were used to prepare cDNAs, which were hybridized to the Whole Genome Human 44k Microarray chips (Agilent Technologies). Data analysis was performed using GeneSpring and Ingenuity Pathway Analysis software. Validation of microarray results was performed by real-time quantitative RT-PCR and Western blot analyses. RESULTS. Expression of ∼130 genes was differentially upregulated by >5-fold, whereas ∼60 genes were differentially downregulated by >2-fold in p52-expressing LNCaP cells. Pathway analysis revealed that the upregulated genes belong to functional categories like cell growth and proliferation, cellular movement, cell-to-cell signaling and interaction, cancer, cell cycle, etc., whereas the downregulated genes were represented by functional categories like cell movement, antigen presentation, and cell death. Six of the top upregulated genes including annexin A2, PLAU, RND3, Twist2, VEGFC, and CXCL1 were validated by real-time PCR and Western blot analysis. CONCLUSIONS. This study provides a comprehensive analysis of genes potentially regulated by NF-κB2/p52 in the LNCaP prostate cancer cell line and provides a rationale for the induction of castration-resistant growth by p52 in LNCaP cells.

AB - PURPOSE. Our previous studies showed that NF-κB2/p52 is involved in the castration-resistant growth of the androgen-sensitive LNCaP prostate cancer cells. The role of NF-κB2/p52 in lymphomagenesis has been studied extensively, but its target genes in other cancers remain unknown. In order to identify genes potentially regulated by p52 in prostate cancer cells, we performed a genome-wide microarray analysis of genes differentially up- or down-regulated by the overexpression of p52 by adenoviral-mediated gene delivery in LNCaP cells. EXPERIMENTAL DESIGN. Total RNAs from vector control-infected and Adeno-p52-infected LNCaP cells were used to prepare cDNAs, which were hybridized to the Whole Genome Human 44k Microarray chips (Agilent Technologies). Data analysis was performed using GeneSpring and Ingenuity Pathway Analysis software. Validation of microarray results was performed by real-time quantitative RT-PCR and Western blot analyses. RESULTS. Expression of ∼130 genes was differentially upregulated by >5-fold, whereas ∼60 genes were differentially downregulated by >2-fold in p52-expressing LNCaP cells. Pathway analysis revealed that the upregulated genes belong to functional categories like cell growth and proliferation, cellular movement, cell-to-cell signaling and interaction, cancer, cell cycle, etc., whereas the downregulated genes were represented by functional categories like cell movement, antigen presentation, and cell death. Six of the top upregulated genes including annexin A2, PLAU, RND3, Twist2, VEGFC, and CXCL1 were validated by real-time PCR and Western blot analysis. CONCLUSIONS. This study provides a comprehensive analysis of genes potentially regulated by NF-κB2/p52 in the LNCaP prostate cancer cell line and provides a rationale for the induction of castration-resistant growth by p52 in LNCaP cells.

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