IKKβ/NFκBp65 activated by interleukin-13 targets the autophagy-related genes LC3B and beclin 1 in fibroblasts co-cultured with breast cancer cells

Li,Wen‑Lin; Xiong,Li‑Xia; Shi,Xiao‑Yu; Xiao,Liang; Qi,Guan‑Yun; Meng,Chuang

doi:10.3892/etm.2016.3054

IKKβ/NFκBp65 activated by interleukin-13 targets the autophagy-related genes LC3B and beclin 1 in fibroblasts co-cultured with breast cancer cells

Authors:
- Wen‑Lin Li
- Li‑Xia Xiong
- Xiao‑Yu Shi
- Liang Xiao
- Guan‑Yun Qi
- Chuang Meng
View Affiliations

Affiliations: Key Laboratory of Medical Biology, Nanchang University, Nanchang, Jiangxi 330006, P.R. China, Department of Pathophysiology, Medical College, Nanchang University, Nanchang, Jiangxi 330006, P.R. China, Molecular Laboratory, Jiangxi University of Traditional Chinese Medicine, Nanchang, Jiangxi 330006, P.R. China
Published online on: February 8, 2016 https://doi.org/10.3892/etm.2016.3054
Pages: 1259-1264
Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Interleukin-13 (IL-13), a Th2 cytokine, plays an important role in fibrosis, inflammation, tissue hyperresponsiveness and tumor development. Although studies have demonstrated that IL‑13 exerts its roles through signal transducer and activator of transcription 6 (STAT6) signaling pathway, recent studies have revealed that I kappa B kinase (IKK)/nuclear factor kappa B (NFκB) pathway may also be involved in. The aim of this study was to investigate whether IL-13 delivers signals to IKKβ/NFκBp65 and whether autophagy genes are IL‑13‑induced the activation of NFκBp65 transcriptional targets in fibroblasts of breast tumor stroma. We examined the phosphorylation of IKKβ, the activation of NFκBp65 and NFκBp65‑targeted autophagy genes in fibroblasts co‑cultured with breast cancer cells under the condition of IL‑13 stimulation. Results of this study showed that IL‑13 induced IKKβ phosphorylation in the fibroblast line ESF co‑cultured with breast cancer cell line BT474, and subsequently NFκBp65 was activated and aimed at beclin 1 and microtubule‑associated protein 1 light chain 3 B (MAP1LC3B or LC3B) in these ESF cells. BMS345541, an inhibitor of IKK/NFκB pathway, significantly inhibited the IL‑13‑induced the activation of NFκB and also inhibited NFκB‑targeted beclin 1 and LC3B expression. Our results suggest that IL-13 regulates beclin 1 and LC3B expression through IKKβ/NFκBp65 in fibroblasts co‑cultured with breast cancer cells, and IL-13 plays role in activating IKKβ/NFκBp65.

View Figures

View References

1	Cirri P and Chiarugi P: Cancer associated fibroblasts: The dark side of the coin. Am J Cancer Res. 1:482–497. 2011.PubMed/NCBI
2	Beacham DA and Cukierman E: Stromagenesis: The changing face of fibroblastic microenvironments during tumor progression. Semin Cancer Biol. 15:329–341. 2005. View Article : Google Scholar : PubMed/NCBI
3	Kalluri R and Zeisberg M: Fibroblasts in cancer. Nat Rev Cancer. 6:392–401. 2006. View Article : Google Scholar : PubMed/NCBI
4	Gabitass RF, Annels NE, Stocken DD, Pandha HA and Middleton GW: Elevated myeloid-derived suppressor cells in pancreatic, esophageal and gastric cancer are an independent prognostic factor and are associated with significant elevation of the Th2 cytokine interleukin-13. Cancer Immunol Immunother. 60:1419–1430. 2011. View Article : Google Scholar : PubMed/NCBI
5	Aspord C, Pedroza-Gonzalez A, Gallegos M, Tindle S, Burton EC, Su D, Marches F, Banchereau J and Palucka AK: Breast cancer instructs dendritic cells to prime interleukin 13-secreting CD4+ T cells that facilitate tumor development. J Exp Med. 204:1037–1047. 2007. View Article : Google Scholar : PubMed/NCBI
6	Chawey C, Bibeau F, Gourgou-Bourgade S, Burlinchon S, Boissiere F, Laune D, Roques S and Lazennec G: Oestrogen receptor negative breast cancers exhibit high cytokine content. Breast Cancer Res. 9:R152007. View Article : Google Scholar : PubMed/NCBI
7	Srabovici N, Mujagic Z, Mujanovic-Mustedanagic J, Muminovic Z, Softic A and Begic L: Interleukin 13 expression in the primary breast cancer tumour tissue. Biochem Med (Zagreb). 21:131–138. 2011. View Article : Google Scholar : PubMed/NCBI
8	Pedroza-Gonzalez A, Xu K, Wu TC, Aspord C, Tindle S, Marches F, Gallegos M, Burton EC, Savino D, Hori T, et al: Thymic stromal lymphopoietin fosters human breast tumor growth by promoting type 2 inflammation. J Exp Med. 208:479–490. 2011. View Article : Google Scholar : PubMed/NCBI
9	Pula B, Jethon A, Piotrowska A, Gomulkiewicz A, Owczarek T, Calik J, Wojnar A, Witkiewicz W, Rys J, Ugorski M, et al: Podoplanin expression by cancer-associated fibroblasts predicts poor outcome in invasive ductal breast carcinoma. Histopathology. 59:1249–1260. 2011. View Article : Google Scholar : PubMed/NCBI
10	Aboussekhra A: Role of cancer-associated fibroblasts in breast cancer development and prognosis. Int J Dev Biol. 55:841–849. 2011. View Article : Google Scholar : PubMed/NCBI
11	Esteve JM and Knecht E: Mechanisms of autophagy and apoptosis: Recent developments in breast cancer cells. World J Biol Chem. 2:232–238. 2011. View Article : Google Scholar : PubMed/NCBI
12	Martinez-Outschoorn UE, Whitaker-Menezes D, Pavlides S, Chiavarina B, Bonuccelli G, Casey T, Tsirigos A, Migneco G, Witkiewicz A, Balliet R, et al: The autophagic tumor stroma model of cancer or ‘battery-operated tumor growth’: A simple solution to the autophagy paradox. Cell Cycle. 9:4297–4306. 2010. View Article : Google Scholar : PubMed/NCBI
13	Nivon M, Richet E, Codogno P, Arrigo AP and Kretz-Remy C: Autophagy activation by NFkappaB is essential for cell survival after heat shock. Autophagy. 5:766–783. 2009. View Article : Google Scholar : PubMed/NCBI
14	Cianfanelli V and Cecconi F: Autophagy-dependent NFκB regulation. Cell Cycle. 11:436–437. 2012. View Article : Google Scholar : PubMed/NCBI
15	Goto K, Chiba Y and Misawa M: IL-13 induces translocation of NF-kappa B in cultured human bronchial smooth muscle cells. Cytokine. 46:96–99. 2009. View Article : Google Scholar : PubMed/NCBI
16	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real- time quantitative PCR and the 2⁻ ^ΔΔCt method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
17	Hayden MS and Ghosh S: Signaling to NF- kappaB. Genes Dev. 18:2195–2224. 2006. View Article : Google Scholar
18	Wynn TA: Type 2 cytokines: Mechanisms and therapeutic strategies. Nat Rev Immunol. 15:271–282. 2015. View Article : Google Scholar : PubMed/NCBI
19	Goenka S and Kaplan MH: Transcriptional regulation by STAT6. Immunol Res. 50:87–96. 2011. View Article : Google Scholar : PubMed/NCBI
20	Salabei JK and Hill BG: Autophagic regulation of smooth muscle cell biology. Redox Biol. 4:97–103. 2015. View Article : Google Scholar : PubMed/NCBI
21	Barker HE, Bird D, Lang G and Erler JT: Tumor-secreted LOXL2 activates fibroblasts through FAK signaling. Mol Cancer Res. 11:1425–1436. 2013. View Article : Google Scholar : PubMed/NCBI
22	Wan F and Lenardo MJ: The nuclear signaling of NF-κB: Current knowledge, new insights and future perspectives. Cell Res. 20:24–33. 2010. View Article : Google Scholar : PubMed/NCBI
23	Criollo A, Senovilla L, Authier H, Maiuri MC, Morselli E, Vitale I, Kepp O, Tasdemir E, Galluzzi L, Shen S, et al: IKK connects autophagy to major stress pathways. Autophagy. 6:189–191. 2010. View Article : Google Scholar : PubMed/NCBI
24	Niso-Santano M, Criollo A, Malik SA, Michaud M, Morselli E, Mariño G, Lachkar S, Galluzzi L, Maiuri MC and Kroemer G: Direct molecular interactions between Beclin1 and the canonical NFκB activation pathway. Autophagy. 8:268–270. 2012. View Article : Google Scholar : PubMed/NCBI
25	Zhang JH: Teaching the basics of autophagy and mitophagy to redox biologists-mechanisms and experimental approaches. Redox Biol. 4:242–259. 2015. View Article : Google Scholar : PubMed/NCBI
26	Kang R, Zeh HJ, Lotze MT and Tang D: The Beclin1 network regulates autophagy and apoptosis. Cell Death Differ. 18:571–580. 2011. View Article : Google Scholar : PubMed/NCBI
27	Copetti T, Bertoli C, Dalla E, Demarchi F and Schneider C: p65/RelA modulates BECN1 transcription and autophagy. Mol Cell Biol. 29:2594–2608. 2009. View Article : Google Scholar : PubMed/NCBI