Beclin 1, LC3 and P62 Expression in Equine Sarcoids.
Abstract: It is well known that δ-bovine papillomaviruses (BPV-1, BPV-2 and BPV-13) are one of the major causative agents of equine sarcoids, the most common equine skin tumors. Different viruses, including papillomaviruses, evolved ingenious strategies to modulate autophagy, a complex process involved in degradation and recycling of old and damaged material. Methods: The aim of this study was to evaluate, by immunohistochemistry (IHC) and Western blot (WB) analysis, the expression of the main related autophagy proteins (Beclin 1, protein light chain 3 (LC3) and P62), in 35 BPV1/2 positive equine sarcoids and 5 BPV negative normal skin samples. Results: Sarcoid samples showed from strong-to-moderate cytoplasmic immunostaining, respectively, for Beclin 1 and P62 in >60% of neoplastic fibroblasts, while LC3 immunostaining was weak to moderate in ≤60% of neoplastic fibroblasts. Western blot analysis confirmed the specificity of the antibodies and revealed no activation of autophagic flux despite Beclin 1 overexpression in sarcoid samples. Conclusions: Results could suggest the activation of the initial phase of autophagy in equine sarcoids, and its impairment during the following steps. The impairment of autophagy could lead to a selection of a quiescent population of fibroblasts, which survive longer in a hypoxic microenvironment and produced more and/or altered collagen.
Publication Date: 2021-12-23 PubMed ID: 35011126PubMed Central: PMC8749869DOI: 10.3390/ani12010020Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
The research focused on studying the expression of key autophagy proteins, namely Beclin 1, LC3 and P62, in horse skin tumors, or equine sarcoids, caused by δ-bovine papillomaviruses. Their findings suggest that initial stages of autophagy may be activated in these tumors, but are impeded in later stages, which could lead to a survival advantage for certain fibroblasts in a low-oxygen environment and influence collagen production.
Research Objectives
- The aim of the study was to observe the expression of Beclin 1, protein light chain 3 (LC3), and P62 proteins in horse skin tumors caused by δ-bovine papillomaviruses (BPV-1, BPV-2 and BPV-13), considered to be one of the major causes of these tumors.
Methods
- The researchers used immunohistochemistry (a technique for detecting proteins in cells of a tissue section) and Western blot analysis, a common method for detection and quantification of individual proteins in a sample.
- They selected 35 BPV1/2 positive equine sarcoid samples and 5 BPV negative normal skin samples for comparison.
Results
- The sarcoid samples demonstrated significant to moderate cytoplasmic immunostaining (an indication of the presence of the target protein) for Beclin 1 and P62 in over 60% of the neoplastic (abnormal growth) fibroblasts, or cells that produce the structural framework of animal tissues.
- LC3 immunostaining was found to be weak to moderate in fewer or equal to 60% of the neoplastic fibroblasts.
- Western blot analysis confirmed the specificity of the antibodies used in the immunohistochemistry and revealed that despite the overexpression of Beclin 1, there was no activation of the autophagic flux, a process which is critical for cellular waste disposal and recycling.
Conclusions
- The findings indicate a possible activation of the initial phase of autophagy in horse skin tumors and suggest impairment during subsequent steps.
- This impaired autophagy process could potentially create a population of fibroblasts that survive longer in a low-oxygen microenvironment and consequently produce more collagen, a key structural protein in skin tissue, or alter its production in some way.
Cite This Article
APA
Martano M, Altamura G, Power K, Liguori P, Restucci B, Borzacchiello G, Maiolino P.
(2021).
Beclin 1, LC3 and P62 Expression in Equine Sarcoids.
Animals (Basel), 12(1), 20.
https://doi.org/10.3390/ani12010020 Publication
Researcher Affiliations
- Department of Veterinary Medicine and Animal Productions, University of Naples Federico II, Via F. Delpino 1, 80137 Naples, Italy.
- Department of Veterinary Medicine and Animal Productions, University of Naples Federico II, Via F. Delpino 1, 80137 Naples, Italy.
- Department of Veterinary Medicine and Animal Productions, University of Naples Federico II, Via F. Delpino 1, 80137 Naples, Italy.
- DVM, EQUINE CLINIC Punto Verde, 81110 Caserta, Italy.
- Department of Veterinary Medicine and Animal Productions, University of Naples Federico II, Via F. Delpino 1, 80137 Naples, Italy.
- Department of Veterinary Medicine and Animal Productions, University of Naples Federico II, Via F. Delpino 1, 80137 Naples, Italy.
- Department of Veterinary Medicine and Animal Productions, University of Naples Federico II, Via F. Delpino 1, 80137 Naples, Italy.
Conflict of Interest Statement
The authors declare no conflict of interest.
References
This article includes 57 references
- Rothman S. How is the balance between protein synthesis and degradation achieved?. Theor Biol Med Model 2010 Jun 23;7:25.
- Klionsky DJ, Emr SD. Autophagy as a regulated pathway of cellular degradation.. Science 2000 Dec 1;290(5497):1717-21.
- Kroemer G. Autophagy: a druggable process that is deregulated in aging and human disease.. J Clin Invest 2015 Jan;125(1):1-4.
- Dikic I. Proteasomal and Autophagic Degradation Systems.. Annu Rev Biochem 2017 Jun 20;86:193-224.
- Parzych KR, Klionsky DJ. An overview of autophagy: morphology, mechanism, and regulation.. Antioxid Redox Signal 2014 Jan 20;20(3):460-73.
- Tooze SA, Schiavo G. Liaisons dangereuses: autophagy, neuronal survival and neurodegeneration.. Curr Opin Neurobiol 2008 Oct;18(5):504-15.
- Münz C. Enhancing immunity through autophagy.. Annu Rev Immunol 2009;27:423-49.
- Tukaj C. The significance of macroautophagy in health and disease.. Folia Morphol (Warsz) 2013 May;72(2):87-93.
- Itakura E, Kishi C, Inoue K, Mizushima N. Beclin 1 forms two distinct phosphatidylinositol 3-kinase complexes with mammalian Atg14 and UVRAG.. Mol Biol Cell 2008 Dec;19(12):5360-72.
- Kabeya Y, Mizushima N, Ueno T, Yamamoto A, Kirisako T, Noda T, Kominami E, Ohsumi Y, Yoshimori T. LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing.. EMBO J 2000 Nov 1;19(21):5720-8.
- Pankiv S, Clausen TH, Lamark T, Brech A, Bruun JA, Outzen H, Øvervatn A, Bjørkøy G, Johansen T. p62/SQSTM1 binds directly to Atg8/LC3 to facilitate degradation of ubiquitinated protein aggregates by autophagy.. J Biol Chem 2007 Aug 17;282(33):24131-45.
- Lavandero S, Chiong M, Rothermel BA, Hill JA. Autophagy in cardiovascular biology.. J Clin Invest 2015 Jan;125(1):55-64.
- Frake RA, Ricketts T, Menzies FM, Rubinsztein DC. Autophagy and neurodegeneration.. J Clin Invest 2015 Jan;125(1):65-74.
- White E. The role for autophagy in cancer.. J Clin Invest 2015 Jan;125(1):42-6.
- Jackson WT. Viruses and the autophagy pathway.. Virology 2015 May;479-480:450-6.
- Mattoscio D, Medda A, Chiocca S. Human Papilloma Virus and Autophagy.. Int J Mol Sci 2018 Jun 15;19(6).
- Roperto S, Russo V, De Falco F, Urraro C, Maiolino P, Del Piero F, Roperto F. Bovine papillomavirus E5 oncoprotein expression and its association with an interactor network in aggresome-autophagy pathway.. Vet Microbiol 2019 Jun;233:39-46.
- Roperto S, Russo V, Rosati A, Ceccarelli DM, Munday JS, Turco MC, Roperto F. Chaperone-assisted selective autophagy in healthy and papillomavirus-associated neoplastic urothelium of cattle.. Vet Microbiol 2018 Jul;221:134-142.
- Borzacchiello G, Roperto F. Bovine papillomaviruses, papillomas and cancer in cattle.. Vet Res 2008 Sep-Oct;39(5):45.
- Lunardi M, de Alcântara BK, Otonel RA, Rodrigues WB, Alfieri AF, Alfieri AA. Bovine papillomavirus type 13 DNA in equine sarcoids.. J Clin Microbiol 2013 Jul;51(7):2167-71.
- Chambers G, Ellsmore VA, O'Brien PM, Reid SWJ, Love S, Campo MS, Nasir L. Association of bovine papillomavirus with the equine sarcoid.. J Gen Virol 2003 May;84(Pt 5):1055-1062.
- Bogaert L, Martens A, Kast WM, Van Marck E, De Cock H. Bovine papillomavirus DNA can be detected in keratinocytes of equine sarcoid tumors.. Vet Microbiol 2010 Dec 15;146(3-4):269-75.
- Marti E, Lazary S, Antczak DF, Gerber H. Report of the first international workshop on equine sarcoid.. Equine Vet J 1993 Sep;25(5):397-407.
- Ragland WL, Keown GH, Spencer GR. Equine sarcoid. Equine Vet. J. 1970;2:2–11.
- Borzacchiello G, Corteggio A. Equine sarcoid: State of the art. Ippologia 2009;20:7–14.
- Nasir L, Campo MS. Bovine papillomaviruses: their role in the aetiology of cutaneous tumours of bovids and equids.. Vet Dermatol 2008 Oct;19(5):243-54.
- Nasir L, Brandt S. Papillomavirus associated diseases of the horse.. Vet Microbiol 2013 Nov 29;167(1-2):159-67.
- Knottenbelt DC. A suggested clinical classification of the equine sarcoid. Clinical techniques in equine. Practice 2005;4:278–295.
- Cochrane CA. Models in vivo of wound healing in the horse and the role of growth factors.. Vet Dermatol 1997 Dec;8(4):259-272.
- Hanson RR. Complications of equine wound management and dermatologic surgery.. Vet Clin North Am Equine Pract 2008 Dec;24(3):663-96, ix.
- Martano M, Corteggio A, Restucci B, De Biase ME, Borzacchiello G, Maiolino P. Extracellular matrix remodeling in equine sarcoid: an immunohistochemical and molecular study.. BMC Vet Res 2016 Feb 2;12:24.
- Martano M, Power K, Restucci B, Pagano I, Altamura G, Borzacchiello G, Maiolino P. Expression of vascular endothelial growth factor (VEGF) in equine sarcoid.. BMC Vet Res 2018 Sep 3;14(1):266.
- Martano M, Altamura G, Power K, Restucci B, Carella F, Borzacchiello G, Maiolino P. Evaluation of Hypoxia-Inducible Factor-1 Alpha (HIF-1α) in Equine Sarcoid: An Immunohistochemical and Biochemical Study.. Pathogens 2020 Jan 14;9(1).
- Yun CW, Lee SH. The Roles of Autophagy in Cancer.. Int J Mol Sci 2018 Nov 5;19(11).
- Borzacchiello G, Russo V, Della Salda L, Roperto S, Roperto F. Expression of platelet-derived growth factor-beta receptor and bovine papillomavirus E5 and E7 oncoproteins in equine sarcoid.. J Comp Pathol 2008 Nov;139(4):231-7.
- Ming He J, Liu PY, Wang J. MicroRNA-17-5p regulates the growth, migration and invasion of the human osteosarcoma cells by modulating the expression of PTEN.. J BUON 2020 Mar-Apr;25(2):1028-1034.
- Katsuragi Y, Ichimura Y, Komatsu M. p62/SQSTM1 functions as a signaling hub and an autophagy adaptor.. FEBS J 2015 Dec;282(24):4672-8.
- Liu JL, Chen FF, Lung J, Lo CH, Lee FH, Lu YC, Hung CH. Prognostic significance of p62/SQSTM1 subcellular localization and LC3B in oral squamous cell carcinoma.. Br J Cancer 2014 Aug 26;111(5):944-54.
- Vega-Rubín-de-Celis S. The Role of Beclin 1-Dependent Autophagy in Cancer.. Biology (Basel) 2019 Dec 22;9(1).
- Won KY, Kim GY, Kim YW, Song JY, Lim SJ. Clinicopathologic correlation of beclin-1 and bcl-2 expression in human breast cancer.. Hum Pathol 2010 Jan;41(1):107-12.
- Jiang ZF, Shao LJ, Wang WM, Yan XB, Liu RY. Decreased expression of Beclin-1 and LC3 in human lung cancer.. Mol Biol Rep 2012 Jan;39(1):259-67.
- Won KY, Kim GY, Lim SJ, Kim YW. Decreased Beclin-1 expression is correlated with the growth of the primary tumor in patients with squamous cell carcinoma and adenocarcinoma of the lung.. Hum Pathol 2012 Jan;43(1):62-8.
- Ding ZB, Shi YH, Zhou J, Qiu SJ, Xu Y, Dai Z, Shi GM, Wang XY, Ke AW, Wu B, Fan J. Association of autophagy defect with a malignant phenotype and poor prognosis of hepatocellular carcinoma.. Cancer Res 2008 Nov 15;68(22):9167-75.
- Miracco C, Cevenini G, Franchi A, Luzi P, Cosci E, Mourmouras V, Monciatti I, Mannucci S, Biagioli M, Toscano M, Moretti D, Lio R, Massi D. Beclin 1 and LC3 autophagic gene expression in cutaneous melanocytic lesions.. Hum Pathol 2010 Apr;41(4):503-12.
- Ahn CH, Jeong EG, Lee JW, Kim MS, Kim SH, Kim SS, Yoo NJ, Lee SH. Expression of beclin-1, an autophagy-related protein, in gastric and colorectal cancers.. APMIS 2007 Dec;115(12):1344-9.
- Kim HS, Lee SH, Do SI, Lim SJ, Park YK, Kim YW. Clinicopathologic correlation of beclin-1 expression in pancreatic ductal adenocarcinoma.. Pathol Res Pract 2011 Apr 15;207(4):247-52.
- Uchida K. Pathologic Changes and Autophagy: New Insights for the Pathogenesis of Animal Diseases.. Vet Pathol 2017 Nov;54(6):881-884.
- Liu JL, Chang KC, Lo CC, Chu PY, Liu CH. Expression of autophagy-related protein Beclin-1 in malignant canine mammary tumors.. BMC Vet Res 2013 Apr 11;9:75.
- Akinduro O, Sully K, Patel A, Robinson DJ, Chikh A, McPhail G, Braun KM, Philpott MP, Harwood CA, Byrne C, O'Shaughnessy RFL, Bergamaschi D. Constitutive Autophagy and Nucleophagy during Epidermal Differentiation.. J Invest Dermatol 2016 Jul;136(7):1460-1470.
- Lee SJ, Kim HP, Jin Y, Choi AM, Ryter SW. Beclin 1 deficiency is associated with increased hypoxia-induced angiogenesis.. Autophagy 2011 Aug;7(8):829-39.
- Tan Q, Wang M, Yu M, Zhang J, Bristow RG, Hill RP, Tannock IF. Role of Autophagy as a Survival Mechanism for Hypoxic Cells in Tumors.. Neoplasia 2016 Jun;18(6):347-55.
- Chen Y, Lu Y, Lu C, Zhang L. Beclin-1 expression is a predictor of clinical outcome in patients with esophageal squamous cell carcinoma and correlated to hypoxia-inducible factor (HIF)-1alpha expression.. Pathol Oncol Res 2009 Sep;15(3):487-93.
- Ranaghan MJ, Durney MA, Mesleh MF, McCarren PR, Garvie CW, Daniels DS, Carey KL, Skepner AP, Levine B, Perez JR. The Autophagy-Related Beclin-1 Protein Requires the Coiled-Coil and BARA Domains To Form a Homodimer with Submicromolar Affinity.. Biochemistry 2017 Dec 26;56(51):6639-6651.
- Del Principe D, Vona R, Giordani L, Straface E, Giammarioli AM. Defective autophagy in fibroblasts may contribute to fibrogenesis in autoimmune processes.. Curr Pharm Des 2011 Dec;17(35):3878-87.
- Martinez-Outschoorn UE, Trimmer C, Lin Z, Whitaker-Menezes D, Chiavarina B, Zhou J, Wang C, Pavlides S, Martinez-Cantarin MP, Capozza F, Witkiewicz AK, Flomenberg N, Howell A, Pestell RG, Caro J, Lisanti MP, Sotgia F. Autophagy in cancer associated fibroblasts promotes tumor cell survival: Role of hypoxia, HIF1 induction and NFκB activation in the tumor stromal microenvironment.. Cell Cycle 2010 Sep 1;9(17):3515-33.
- Everts V, van der Zee E, Creemers L, Beertsen W. Phagocytosis and intracellular digestion of collagen, its role in turnover and remodelling.. Histochem J 1996 Apr;28(4):229-45.
- Wang ZH, Xu L, Wang Y, Cao MQ, Li L, Bai T. Clinicopathologic correlations between human papillomavirus 16 infection and Beclin 1 expression in human cervical cancer.. Int J Gynecol Pathol 2011 Jul;30(4):400-6.
Citations
This article has been cited 3 times.- Power K, Martano M, Piscopo N, Viola P, Altamura G, Veneziano V, Carvajal Urueña A, Esposito L. Prevalence of Eucoleus garfiai in Wild Boars Hunted at Different Altitudes in the Campania and Latium Regions (Italy). Animals (Basel) 2023 Feb 17;13(4).
- Lu F, Zhong J, Zhou Q, Yu Y, Liang M, Yuan Y, Xie A, Cheng J, Shu P, Hao J. Okanin Attenuates Mitochondrial Dysfunction and Apoptosis in UVA-Induced HaCaT Cells by Mitophagy Through SIRT3 Pathway. Antioxidants (Basel) 2025 Aug 23;14(9).
- Cui YN, Tian N, Luo YH, Zhao JJ, Bi CF, Gou Y, Liu J, Feng K, Zhang JF. High-dose Vitamin C injection ameliorates against sepsis-induced myocardial injury by anti-apoptosis, anti-inflammatory and pro-autophagy through regulating MAPK, NF-κB and PI3K/AKT/mTOR signaling pathways in rats. Aging (Albany NY) 2024 Apr 19;16(8):6937-6953.
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists