Cigarette smoke induces C/EBP-β-mediated activation of mir-31 in normal human respiratory epithelia and lung cancer cells

Sichuan Xi, Maocheng Yang, Yongguang Tao, Hong Xu, Jigui Shan, Suzanne Inchauste, Mary Zhang, Leandro Mercedes, Julie A. Hong, Mahadev Rao, David S. Schrump

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Abstract

Background: Limited information is available regarding mechanisms by which MIRNAS contribute to pulmonary carcinogenesis. The present study was undertaken to examine expression and function of MIRNAS induced by cigarette smoke condensate (CSC) in normal human respiratory epithelia and lung cancer cells. Methodology: Micro-array and quantitative RT-PCR (QRT-PCR) techniques were used to assess MIRNA and host gene expression in cultured cells, and surgical specimens. Software-guided analysis, RNA cross-link immunoprecipitation (CLIP), 39 UTR luciferase reporter assays, QRT-PCR, focused super-arrays and western blot techniques were used to identify and confirm targets of miR-31. Chromatin immunoprecipitation (ChIP) techniques were used to evaluate histone marks and transcription factors within the LOC554202 promoter. Cell count and xenograft experiments were used to assess effects of miR-31 on proliferation and tumorigenicity of lung cancer cells. Results: CSC significantly increased miR-31 expression and activated LOC554202 in normal respiratory epithelia and lung cancer cells; miR-31 and LOC554202 expression persisted following discontinuation of CSC exposure. miR-31 and LOC554202 expression levels were significantly elevated in lung cancer specimens relative to adjacent normal lung tissues. CLIP and reporter assays demonstrated direct interaction of miR-31 with Dickkopf-1 (Dkk-1) and DACT-3. Over-expression of miR-31 markedly diminished Dkk-1 and DACT3 expression levels in normal respiratory epithelia and lung cancer cells. Knock-down of miR-31 increased Dkk-1 and DACT3 levels, and abrogated CSC-mediated decreases in Dkk-1 and DACT-3 expression. Furthermore, over-expression of miR-31 diminished SFRP1, SFRP4, and WIF-1, and increased Wnt-5a expression. CSC increased H3K4Me3, H3K9/14Ac and C/EBP-b levels within the LOC554202 promoter. Knock-down of C/EBP-b abrogated CSC-mediated activation of LOC554202. Over-expression of miR-31 significantly enhanced proliferation and tumorigenicity of lung cancer cells; knock-down of miR-31 inhibited growth of these cells. Conclusions:Cigarette smoke induces expression of miR-31 targeting several antagonists of cancer stem cell signaling in normal respiratory epithelia and lung cancer cells. miR-31 functions as an oncomir during human pulmonary carcinogenesis.

Original languageEnglish
Article numbere13764
JournalPLoS One
Volume5
Issue number10
DOIs
Publication statusPublished - 2010

Fingerprint

respiratory mucosa
Respiratory Mucosa
cigarettes
lung neoplasms
smoke
Smoke
Tobacco Products
Lung Neoplasms
Chemical activation
Cells
lungs
Immunoprecipitation
Lung
carcinogenesis
Carcinogenesis
Histone Code
reverse transcriptase polymerase chain reaction
Assays
promoter regions
Untranslated Regions

All Science Journal Classification (ASJC) codes

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Xi, Sichuan ; Yang, Maocheng ; Tao, Yongguang ; Xu, Hong ; Shan, Jigui ; Inchauste, Suzanne ; Zhang, Mary ; Mercedes, Leandro ; Hong, Julie A. ; Rao, Mahadev ; Schrump, David S. / Cigarette smoke induces C/EBP-β-mediated activation of mir-31 in normal human respiratory epithelia and lung cancer cells. In: PLoS One. 2010 ; Vol. 5, No. 10.
@article{b52bc7a1d8b34879b272c5f7aa2e9d50,
title = "Cigarette smoke induces C/EBP-β-mediated activation of mir-31 in normal human respiratory epithelia and lung cancer cells",
abstract = "Background: Limited information is available regarding mechanisms by which MIRNAS contribute to pulmonary carcinogenesis. The present study was undertaken to examine expression and function of MIRNAS induced by cigarette smoke condensate (CSC) in normal human respiratory epithelia and lung cancer cells. Methodology: Micro-array and quantitative RT-PCR (QRT-PCR) techniques were used to assess MIRNA and host gene expression in cultured cells, and surgical specimens. Software-guided analysis, RNA cross-link immunoprecipitation (CLIP), 39 UTR luciferase reporter assays, QRT-PCR, focused super-arrays and western blot techniques were used to identify and confirm targets of miR-31. Chromatin immunoprecipitation (ChIP) techniques were used to evaluate histone marks and transcription factors within the LOC554202 promoter. Cell count and xenograft experiments were used to assess effects of miR-31 on proliferation and tumorigenicity of lung cancer cells. Results: CSC significantly increased miR-31 expression and activated LOC554202 in normal respiratory epithelia and lung cancer cells; miR-31 and LOC554202 expression persisted following discontinuation of CSC exposure. miR-31 and LOC554202 expression levels were significantly elevated in lung cancer specimens relative to adjacent normal lung tissues. CLIP and reporter assays demonstrated direct interaction of miR-31 with Dickkopf-1 (Dkk-1) and DACT-3. Over-expression of miR-31 markedly diminished Dkk-1 and DACT3 expression levels in normal respiratory epithelia and lung cancer cells. Knock-down of miR-31 increased Dkk-1 and DACT3 levels, and abrogated CSC-mediated decreases in Dkk-1 and DACT-3 expression. Furthermore, over-expression of miR-31 diminished SFRP1, SFRP4, and WIF-1, and increased Wnt-5a expression. CSC increased H3K4Me3, H3K9/14Ac and C/EBP-b levels within the LOC554202 promoter. Knock-down of C/EBP-b abrogated CSC-mediated activation of LOC554202. Over-expression of miR-31 significantly enhanced proliferation and tumorigenicity of lung cancer cells; knock-down of miR-31 inhibited growth of these cells. Conclusions:Cigarette smoke induces expression of miR-31 targeting several antagonists of cancer stem cell signaling in normal respiratory epithelia and lung cancer cells. miR-31 functions as an oncomir during human pulmonary carcinogenesis.",
author = "Sichuan Xi and Maocheng Yang and Yongguang Tao and Hong Xu and Jigui Shan and Suzanne Inchauste and Mary Zhang and Leandro Mercedes and Hong, {Julie A.} and Mahadev Rao and Schrump, {David S.}",
year = "2010",
doi = "10.1371/journal.pone.0013764",
language = "English",
volume = "5",
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Xi, S, Yang, M, Tao, Y, Xu, H, Shan, J, Inchauste, S, Zhang, M, Mercedes, L, Hong, JA, Rao, M & Schrump, DS 2010, 'Cigarette smoke induces C/EBP-β-mediated activation of mir-31 in normal human respiratory epithelia and lung cancer cells', PLoS One, vol. 5, no. 10, e13764. https://doi.org/10.1371/journal.pone.0013764

Cigarette smoke induces C/EBP-β-mediated activation of mir-31 in normal human respiratory epithelia and lung cancer cells. / Xi, Sichuan; Yang, Maocheng; Tao, Yongguang; Xu, Hong; Shan, Jigui; Inchauste, Suzanne; Zhang, Mary; Mercedes, Leandro; Hong, Julie A.; Rao, Mahadev; Schrump, David S.

In: PLoS One, Vol. 5, No. 10, e13764, 2010.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Cigarette smoke induces C/EBP-β-mediated activation of mir-31 in normal human respiratory epithelia and lung cancer cells

AU - Xi, Sichuan

AU - Yang, Maocheng

AU - Tao, Yongguang

AU - Xu, Hong

AU - Shan, Jigui

AU - Inchauste, Suzanne

AU - Zhang, Mary

AU - Mercedes, Leandro

AU - Hong, Julie A.

AU - Rao, Mahadev

AU - Schrump, David S.

PY - 2010

Y1 - 2010

N2 - Background: Limited information is available regarding mechanisms by which MIRNAS contribute to pulmonary carcinogenesis. The present study was undertaken to examine expression and function of MIRNAS induced by cigarette smoke condensate (CSC) in normal human respiratory epithelia and lung cancer cells. Methodology: Micro-array and quantitative RT-PCR (QRT-PCR) techniques were used to assess MIRNA and host gene expression in cultured cells, and surgical specimens. Software-guided analysis, RNA cross-link immunoprecipitation (CLIP), 39 UTR luciferase reporter assays, QRT-PCR, focused super-arrays and western blot techniques were used to identify and confirm targets of miR-31. Chromatin immunoprecipitation (ChIP) techniques were used to evaluate histone marks and transcription factors within the LOC554202 promoter. Cell count and xenograft experiments were used to assess effects of miR-31 on proliferation and tumorigenicity of lung cancer cells. Results: CSC significantly increased miR-31 expression and activated LOC554202 in normal respiratory epithelia and lung cancer cells; miR-31 and LOC554202 expression persisted following discontinuation of CSC exposure. miR-31 and LOC554202 expression levels were significantly elevated in lung cancer specimens relative to adjacent normal lung tissues. CLIP and reporter assays demonstrated direct interaction of miR-31 with Dickkopf-1 (Dkk-1) and DACT-3. Over-expression of miR-31 markedly diminished Dkk-1 and DACT3 expression levels in normal respiratory epithelia and lung cancer cells. Knock-down of miR-31 increased Dkk-1 and DACT3 levels, and abrogated CSC-mediated decreases in Dkk-1 and DACT-3 expression. Furthermore, over-expression of miR-31 diminished SFRP1, SFRP4, and WIF-1, and increased Wnt-5a expression. CSC increased H3K4Me3, H3K9/14Ac and C/EBP-b levels within the LOC554202 promoter. Knock-down of C/EBP-b abrogated CSC-mediated activation of LOC554202. Over-expression of miR-31 significantly enhanced proliferation and tumorigenicity of lung cancer cells; knock-down of miR-31 inhibited growth of these cells. Conclusions:Cigarette smoke induces expression of miR-31 targeting several antagonists of cancer stem cell signaling in normal respiratory epithelia and lung cancer cells. miR-31 functions as an oncomir during human pulmonary carcinogenesis.

AB - Background: Limited information is available regarding mechanisms by which MIRNAS contribute to pulmonary carcinogenesis. The present study was undertaken to examine expression and function of MIRNAS induced by cigarette smoke condensate (CSC) in normal human respiratory epithelia and lung cancer cells. Methodology: Micro-array and quantitative RT-PCR (QRT-PCR) techniques were used to assess MIRNA and host gene expression in cultured cells, and surgical specimens. Software-guided analysis, RNA cross-link immunoprecipitation (CLIP), 39 UTR luciferase reporter assays, QRT-PCR, focused super-arrays and western blot techniques were used to identify and confirm targets of miR-31. Chromatin immunoprecipitation (ChIP) techniques were used to evaluate histone marks and transcription factors within the LOC554202 promoter. Cell count and xenograft experiments were used to assess effects of miR-31 on proliferation and tumorigenicity of lung cancer cells. Results: CSC significantly increased miR-31 expression and activated LOC554202 in normal respiratory epithelia and lung cancer cells; miR-31 and LOC554202 expression persisted following discontinuation of CSC exposure. miR-31 and LOC554202 expression levels were significantly elevated in lung cancer specimens relative to adjacent normal lung tissues. CLIP and reporter assays demonstrated direct interaction of miR-31 with Dickkopf-1 (Dkk-1) and DACT-3. Over-expression of miR-31 markedly diminished Dkk-1 and DACT3 expression levels in normal respiratory epithelia and lung cancer cells. Knock-down of miR-31 increased Dkk-1 and DACT3 levels, and abrogated CSC-mediated decreases in Dkk-1 and DACT-3 expression. Furthermore, over-expression of miR-31 diminished SFRP1, SFRP4, and WIF-1, and increased Wnt-5a expression. CSC increased H3K4Me3, H3K9/14Ac and C/EBP-b levels within the LOC554202 promoter. Knock-down of C/EBP-b abrogated CSC-mediated activation of LOC554202. Over-expression of miR-31 significantly enhanced proliferation and tumorigenicity of lung cancer cells; knock-down of miR-31 inhibited growth of these cells. Conclusions:Cigarette smoke induces expression of miR-31 targeting several antagonists of cancer stem cell signaling in normal respiratory epithelia and lung cancer cells. miR-31 functions as an oncomir during human pulmonary carcinogenesis.

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U2 - 10.1371/journal.pone.0013764

DO - 10.1371/journal.pone.0013764

M3 - Article

VL - 5

JO - PLoS One

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