Skip to main content
main-content

18.12.2017 | Original Research Paper | Ausgabe 4/2018

Inflammation Research 4/2018

Methylation of cytokines gene promoters in IL-1β-treated human intestinal epithelial cells

Zeitschrift:
Inflammation Research > Ausgabe 4/2018
Autoren:
Fabio Caradonna, Ilenia Cruciata, Ilaria Schifano, Chiara La Rosa, Flores Naselli, Roberto Chiarelli, Anna Perrone, Carla Gentile
Wichtige Hinweise
Responsible Editor: Yoshiya Tanaka.

Electronic supplementary material

The online version of this article (https://​doi.​org/​10.​1007/​s00011-017-1124-5) contains supplementary material, which is available to authorized users.
Fabio Caradonna and Carla Gentile contributed equally.

Abstract

Objective and design

Epigenetic regulation is important in the activation of inflammatory cells. In the present study, we evaluated if DNA-methylation variations are involved in Interleukin-1β (IL-1β)-induced intestinal epithelial cells activation.

Materials and methods

Differentiated Caco-2 cells were exposed to IL-1β or to 5-azadeoxycytidine (5-azadC) for 24 or 48 h. Genome-wide methylation status was evaluated, while DNA methylation status at the promoter region of the gene encoding interleukin-6, 8 and 10 (IL-6, 8 and 10) was estimated. The levels of the corresponding gene products as well as DNA methyltransferases (DNMTs) quantity were assessed.

Results

IL-1β decreased genomic methylation of human intestinal epithelial cells and induced demethylation at cg-specific sites at the promoter of pro-inflammatory genes IL6 and IL8; conversely it did not change the methylation of the IL10 promoter. IL-1β also increased the release of IL-6 and IL-8 but did not change the IL-10 expression. Finally, cell exposure to IL-1β decreased the DNMT3b expression, increased DNMT3a and was not able to change DNMT1 expression.

Conclusions

Our results suggest a potential role of IL-1β as modulator of DNA methylation in activated differentiated Caco-2 cell line.

Bitte loggen Sie sich ein, um Zugang zu diesem Inhalt zu erhalten

★ PREMIUM-INHALT
e.Med Interdisziplinär

Mit e.Med Interdisziplinär erhalten Sie Zugang zu allen CME-Fortbildungen und Fachzeitschriften auf SpringerMedizin.de. Zusätzlich können Sie eine Zeitschrift Ihrer Wahl in gedruckter Form beziehen – ohne Aufpreis.

Weitere Produktempfehlungen anzeigen
Zusatzmaterial
Fig. 1 supplementary – Representative MSRE-PCR revealing methylation status of IL6/1 cg site in Caco-2 cells treated with IL-1β. Matched pairs of specific PCR products from undigested or digested (Hh) DNA from either untreated or IL-1β-treated Caco-2 cells for 24 h. Densitometry of the bands is showed by histograms which represent the mean of triplicate experiments with similar results. In each panel, values not sharing the same letter were significantly different (p < 0.001, according to chi square test). Hh = HhaI. (TIF 3579 KB)
11_2017_1124_MOESM1_ESM.tif
Fig. 2 supplementary – Representative MSRE-PCR revealing methylation status of IL6/3 cg site in Caco-2 cells treated with IL-1β. Matched pairs of specific PCR products from undigested or digested (H or M) DNA from either untreated or IL-1β-treated Caco-2 cells for 24 h. Densitometry of the bands is showed by histograms which represent the mean of triplicate experiments with similar results. M, H = Msp, HpaII respectively. (TIF 3400 KB)
11_2017_1124_MOESM2_ESM.tif
Fig. 3 supplementary – Representative MSRE-PCR revealing methylation status of IL6/4 cg site in Caco-2 cells treated with IL-1β. Matched pairs of specific PCR products from undigested or digested (Hh) DNA from either untreated or IL-1β-treated Caco-2 cells for 24 h. Densitometry of the bands is showed by histograms which represent the mean of triplicate experiments with similar results. Hh = HhaI. (TIF 3344 KB)
11_2017_1124_MOESM3_ESM.tif
Fig. 4 supplementary – Representative MSRE-PCR revealing methylation status of IL6/6 cg site in Caco-2 cells treated with IL-1β. Matched pairs of specific PCR products from undigested or digested (Hh) DNA from either untreated or IL-1β-treated Caco-2 cells for 24 h. Densitometry of the bands is showed by histograms which represent the mean of triplicate experiments with similar results. In each panel, values not sharing the same letter were significantly different (p < 0.001, according to chi square test). Hh = HhaI. (TIF 3444 KB)
11_2017_1124_MOESM4_ESM.tif
Fig. 5 supplementary – Representative MSRE-PCR revealing methylation status of IL6/7 cg site in Caco-2 cells treated with IL-1β. Matched pairs of specific PCR products from undigested or digested (H or M) DNA from either untreated or IL-1β-treated Caco-2 cells for 24 h. Densitometry of the bands is showed by histograms which represent the mean of triplicate experiments with similar results. In each panel, values not sharing the same letter were significantly different (p < 0.05, according to chi square test). M, H = Msp, HpaII respectively. (TIF 3424 KB)
11_2017_1124_MOESM5_ESM.tif
Fig. 6 supplementary – Representative MSRE-PCR revealing methylation status of IL8/1 cg site in Caco-2 cells treated with IL-1β. Matched pairs of specific PCR products from undigested or digested (H or M) DNA from either untreated or IL-1β-treated Caco-2 cells for 24 h. Densitometry of the bands is showed by histograms which represent the mean of triplicate experiments with similar results. M, H = Msp, HpaII respectively. (TIF 3491 KB)
11_2017_1124_MOESM6_ESM.tif
Fig. 7 supplementary – Representative MSRE-PCR revealing methylation status of IL10/1 cg site in Caco-2 cells treated with IL-1β. Matched pairs of specific PCR products from undigested or digested (H or M) DNA from either untreated or IL-1β-treated Caco-2 cells for 24 h. Densitometry of the bands is showed by histograms which represent the mean of triplicate experiments with similar results. M, H = Msp, HpaII respectively. (TIF 3367 KB)
11_2017_1124_MOESM7_ESM.tif
Fig. 8 supplementary – Representative MSRE-PCR revealing methylation status of IL10/2 cg site in Caco-2 cells treated with IL-1β. Matched pairs of specific PCR products from undigested or digested (Hh) DNA from either untreated or IL-1β-treated Caco-2 cells for 24 h. Densitometry of the bands is showed by histograms which represent the mean of triplicate experiments with similar results. Hh = HhaI. (TIF 3098 KB)
11_2017_1124_MOESM8_ESM.tif
Literatur
Über diesen Artikel

Weitere Artikel der Ausgabe 4/2018

Inflammation Research 4/2018 Zur Ausgabe
  1. Das kostenlose Testabonnement läuft nach 14 Tagen automatisch und formlos aus. Dieses Abonnement kann nur einmal getestet werden.

Neu im Fachgebiet Innere Medizin

Meistgelesene Bücher aus der Inneren Medizin

2017 | Buch

Rheumatologie aus der Praxis

Entzündliche Gelenkerkrankungen – mit Fallbeispielen

Dieses Fachbuch macht mit den wichtigsten chronisch entzündlichen Gelenk- und Wirbelsäulenerkrankungen vertraut. Anhand von über 40 instruktiven Fallbeispielen werden anschaulich diagnostisches Vorgehen, therapeutisches Ansprechen und der Verlauf …

Herausgeber:
Rudolf Puchner

2016 | Buch

Ambulant erworbene Pneumonie

Was, wann, warum – Dieses Buch bietet differenzierte Diagnostik und Therapie der ambulant erworbenen Pneumonie zur sofortigen sicheren Anwendung. Entsprechend der neuesten Studien und Leitlinien aller wichtigen Fachgesellschaften.

Herausgeber:
Santiago Ewig

Mail Icon II Newsletter

Bestellen Sie unseren kostenlosen Newsletter Update Innere Medizin und bleiben Sie gut informiert – ganz bequem per eMail.

© Springer Medizin 

Bildnachweise