{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T04:56:24Z","timestamp":1791262584932,"version":"4.1.0"},"reference-count":35,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2019,3,19]],"date-time":"2019-03-19T00:00:00Z","timestamp":1552953600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cancers"],"abstract":"<jats:p>In neoplastic conditions, autophagy may act as a tumor suppressor avoiding the accumulation of damaged proteins and organelles or as a mechanism of cell survival promoting the tumor growth. Although ultrastructural analysis has been considered the traditional method to identify autophagy, some proteins such as microtubule-associated protein 1 light chain 3 (LC3A\/B), Beclin-1 and activating molecule in Beclin-1-regulated autophagy protein-1 (AMBRA-1) may be considered as markers of autophagy-assisted cancerogenesis. Herein, we analyzed a cohort of advanced tubular gastric adenocarcinomas by the abovementioned immunohistochemical antisera; through immunohistochemistry, autophagy (A-IHC) is diagnosed when at least two out of the three proteins are positive in the samples. Immunostaining for LC3A\/B, Beclin-1, and AMBRA-1 was exclusively found in neoplastic elements, but not in surrounding stromal cells. In detail, LC3A\/B and Beclin 1 were expressed both in the cytoplasm and in the nucleus of the cancer cells, while AMBRA-1 was preferentially localized in the nucleus, mainly in high grade cases. LC3A\/B, Beclin 1, and AMBRA-1 expression were positive in 18 (56.2%), 17 (53.1%), and 12 (37.5%) cases, respectively. The sensibility and specificity of LC3A\/B and Beclin-1 ranged from 81.25% to 93.75%, with high efficiency (90.63%) for Beclin-1. Moreover, the ultrastructural autophagic index (AI) was also available in all cases. All high-grade cases documented a Ki-67 labelling index (LI) \u2265 30%, even if three low-grade cases revealed a high Ki-67 value; p53 positivity was encountered in 21\/32 (65.62%) of cases, independently of the tumor grade. A statistically significant correlation among A-IHC and clinicopathological parameters such as grade, stage, clinical course, Ki-67 LI and AI was revealed. Univariate analysis documented a significant p-value for the same autophagic variables. Additionally, multivariate survival analysis identified the grade, AI and A-IHC as independent significant variables. Finally, the overall survival curves of all cases of gastric tubular adenocarcinoma were greatly dependent on A-IHC. Therefore, we suggest that autophagic-related proteins might be considered promising predictive prognostic factors of advanced gastric cancer. Further investigations may be required to determine whether new targeted therapies should be addressed to autophagy-related proteins.<\/jats:p>","DOI":"10.3390\/cancers11030389","type":"journal-article","created":{"date-parts":[[2019,3,19]],"date-time":"2019-03-19T12:12:25Z","timestamp":1552997545000},"page":"389","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":19,"title":["Immunohistochemical Expression of Autophagy-Related Proteins in Advanced Tubular Gastric Adenocarcinomas and Its Implications"],"prefix":"10.3390","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0493-7621","authenticated-orcid":false,"given":"Antonio","family":"Ieni","sequence":"first","affiliation":[{"name":"Department of Human Pathology in Adult and Developmental Age \u201cGaetano Barresi\u201d, Section of Pathology, University of Messina, 98123 Messina, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Roberta","family":"Cardia","sequence":"additional","affiliation":[{"name":"Department of Human Pathology in Adult and Developmental Age \u201cGaetano Barresi\u201d, Section of Pathology, University of Messina, 98123 Messina, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giuseppe","family":"Giuffr\u00e8","sequence":"additional","affiliation":[{"name":"Department of Human Pathology in Adult and Developmental Age \u201cGaetano Barresi\u201d, Section of Pathology, University of Messina, 98123 Messina, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luciana","family":"Rigoli","sequence":"additional","affiliation":[{"name":"Department of Human Pathology in Adult and Developmental Age \u201cGaetano Barresi\u201d, Section of Pathology, University of Messina, 98123 Messina, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rosario Alberto","family":"Caruso","sequence":"additional","affiliation":[{"name":"Department of Human Pathology in Adult and Developmental Age \u201cGaetano Barresi\u201d, Section of Pathology, University of Messina, 98123 Messina, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giovanni","family":"Tuccari","sequence":"additional","affiliation":[{"name":"Department of Human Pathology in Adult and Developmental Age \u201cGaetano Barresi\u201d, Section of Pathology, University of Messina, 98123 Messina, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2019,3,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"313","DOI":"10.1016\/j.cell.2010.01.028","article-title":"Methods in mammalian autophagy research","volume":"140","author":"Mizushima","year":"2010","journal-title":"Cell"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1717","DOI":"10.1126\/science.290.5497.1717","article-title":"Autophagy as a regulated pathway of cellular degradation","volume":"290","author":"Klionsky","year":"2000","journal-title":"Science"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"445","DOI":"10.4161\/auto.19496","article-title":"Guidelines for the use and interpretation of assays for monitoring autophagy","volume":"8","author":"Klionsky","year":"2012","journal-title":"Autophagy"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Bento, C.F., Ashkenazi, A., Jimenez-Sanchez, M., and Rubinsztein, D.C. (2016). The Parkinson\u2019s disease-associated genes ATP13A2 and SYT11 regulate autophagy via a common pathway. Nat. Commun., 7.","DOI":"10.1038\/ncomms11803"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1803","DOI":"10.2147\/OTT.S132508","article-title":"Understanding the role of PD-L1\/PD1 pathway blockade and autophagy in cancer therapy","volume":"10","author":"Robainas","year":"2017","journal-title":"Oncol. Targets Ther."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"708","DOI":"10.1002\/path.5222","article-title":"Autophagy, Cancer Stem Cells & Drug Resistance","volume":"247","author":"Smith","year":"2018","journal-title":"J. Pathol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1080\/01913123.2017.1388322","article-title":"Autophagy in advanced low- and high-grade tubular adenocarcinomas of the stomach: An ultrastructural investigation","volume":"42","author":"Caruso","year":"2018","journal-title":"Ultrastruct. Pathol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1016\/j.cca.2018.11.028","article-title":"Autophagy and its role in gastric cancer","volume":"489","author":"Cao","year":"2019","journal-title":"Clin. Chim. Acta"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1387","DOI":"10.4161\/auto.7.11.17348","article-title":"Spatio-temporal association between mTOR and autophagy during cellular senescence","volume":"7","author":"Young","year":"2011","journal-title":"Autophagy"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"147","DOI":"10.1016\/j.ymeth.2016.05.022","article-title":"Quantifying autophagy: Measuring LC3 puncta and autolysosome formation in cells using multispectral imaging flow cytometry","volume":"112","author":"Pugsley","year":"2017","journal-title":"Methods"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"726","DOI":"10.1038\/nrc1692","article-title":"The role of autophagy in cancer development and response to therapy","volume":"5","author":"Kondo","year":"2005","journal-title":"Nat. Rev. Cancer"},{"key":"ref_12","first-page":"129","article-title":"Clinicopathological Correlations of Autophagy-related Proteins LC3, Beclin 1 and p62 in Gastric Cancer","volume":"36","author":"Masuda","year":"2016","journal-title":"Anticancer Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1038\/ncb3072","article-title":"AMBRA1 links autophagy to cell proliferation and tumorigenesis by promoting c-Myc dephosphorylation and degradation","volume":"17","author":"Cianfanelli","year":"2015","journal-title":"Nat. Cell Biol."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Chude, C.I., and Amaravadi, R.K. (2017). Targeting Autophagy in Cancer: Update on Clinical Trials and Novel Inhibitors. Int. J. Mol. Sci., 18.","DOI":"10.3390\/ijms18061279"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1016\/j.ccr.2006.06.001","article-title":"Autophagy promotes tumor cell survival and restricts necrosis, inflammation, and tumorigenesis","volume":"10","author":"Degenhardt","year":"2006","journal-title":"Cancer Cell"},{"key":"ref_16","first-page":"3831","article-title":"Prognostic value of autophagy related proteins ULK1, Beclin 1, ATG3, ATG5, ATG7, ATG9, ATG10, ATG12, LC3B and p62\/SQSTM1 in gastric cancer","volume":"8","author":"Cao","year":"2016","journal-title":"Am. J. Transl. Res."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1","DOI":"10.3233\/CBM-160613","article-title":"Autophagy regulation and its role in gastric cancer and colorectal cancer","volume":"17","author":"Zhou","year":"2016","journal-title":"Cancer Biomark. Sect. Dis. Mark."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"189","DOI":"10.1186\/s12957-016-0946-x","article-title":"Prognostic relevance of autophagy-related markers LC3, p62\/sequestosome 1, Beclin-1 and ULK1 in colorectal cancer patients with respect to KRAS mutational status","volume":"14","author":"Schmitz","year":"2016","journal-title":"World J. Surg. Oncol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"7084","DOI":"10.18632\/oncotarget.3054","article-title":"Autophagy-based survival prognosis in human colorectal carcinoma","volume":"6","author":"Yang","year":"2015","journal-title":"Oncotarget"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"4779","DOI":"10.3748\/wjg.v17.i43.4779","article-title":"Autophagy-related proteins Beclin-1 and LC3 predict cetuximab efficacy in advanced colorectal cancer","volume":"17","author":"Guo","year":"2011","journal-title":"World J. Gastroenterol."},{"key":"ref_21","first-page":"3882","article-title":"Expression and clinical significances of Beclin1, LC3 and mTOR in colorectal cancer","volume":"8","author":"Wu","year":"2015","journal-title":"Int. J. Clin. Exp. Pathol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"370","DOI":"10.1158\/1078-0432.CCR-11-1282","article-title":"Punctate LC3B expression is a common feature of solid tumors and associated with proliferation, metastasis, and poor outcome","volume":"18","author":"Lazova","year":"2012","journal-title":"Clin. Cancer Res. J. Am. Assoc. Cancer Res."},{"key":"ref_23","first-page":"723176","article-title":"Correlation of altered expression of the autophagy marker lc3b with poor prognosis in astrocytoma","volume":"2014","author":"Winardi","year":"2014","journal-title":"Bio. Med. Res. Int."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"El-Mashed, S., O\u2019Donovan, T.R., Kay, E.W., Abdallah, A.R., Cathcart, M.-C., O\u2019Sullivan, J., O\u2019Grady, A., Reynolds, J., O\u2019Reilly, S., and O\u2019Sullivan, G.C. (2015). LC3B globular structures correlate with survival in esophageal adenocarcinoma. BMC Cancer, 15.","DOI":"10.1186\/s12885-015-1574-5"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"944","DOI":"10.1038\/bjc.2014.355","article-title":"Prognostic significance of p62\/SQSTM1 subcellular localization and LC3B in oral squamous cell carcinoma","volume":"111","author":"Liu","year":"2014","journal-title":"Br. J. Cancer"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"12225","DOI":"10.1007\/s13277-014-2531-7","article-title":"Autophagic LC3B overexpression correlates with malignant progression and predicts a poor prognosis in hepatocellular carcinoma","volume":"35","author":"Wu","year":"2014","journal-title":"Tumour Biol. J. Int. Soc. Oncodev. Biol. Med."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"839","DOI":"10.1038\/cr.2007.78","article-title":"Physiological functions of Atg6\/Beclin 1: A unique autophagy-related protein","volume":"17","author":"Cao","year":"2007","journal-title":"Cell Res."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Bortnik, S., and Gorski, S.M. (2017). Clinical Applications of Autophagy Proteins in Cancer: From Potential Targets to Biomarkers. Int. J. Mol. Sci, 18.","DOI":"10.3390\/ijms18071496"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1209","DOI":"10.1038\/sj.bjc.6605904","article-title":"Beclin 1 over-and underexpression in colorectal cancer: Distinct patterns relate to prognosis and tumour hypoxia","volume":"103","author":"Koukourakis","year":"2010","journal-title":"Br. J. Cancer"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"7317","DOI":"10.1007\/s13277-014-2060-4","article-title":"The prognostic value of autophagy-related markers beclin-1 and microtubule-associated protein light chain 3B in cancers: A systematic review and meta-analysis","volume":"35","author":"He","year":"2014","journal-title":"Tumour Biol. J. Int. Soc. Oncodev. Biol. Med."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"829","DOI":"10.1097\/MPA.0b013e318279d0dc","article-title":"Prognostic significance of autophagy-related protein expression in resected pancreatic ductal adenocarcinoma","volume":"42","author":"Ko","year":"2013","journal-title":"Pancreas"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1007\/s11596-017-1691-2","article-title":"Prognostic significance of autophagy-related proteins expression in resected human gastric adenocarcinoma","volume":"37","author":"Qu","year":"2017","journal-title":"J. Huazhong Univ. Sci. Technol. Med. Sci."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"3478","DOI":"10.1158\/1078-0432.CCR-10-2372","article-title":"Measurements of tumor cell autophagy predict invasiveness, resistance to chemotherapy, and survival in melanoma","volume":"17","author":"Ma","year":"2011","journal-title":"Clin. Cancer Res."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Schl\u00e4fli, A.M., Berezowska, S., Adams, O., Langer, R., and Tschan, M.P. (2015). Reliable LC3 and p62 autophagy marker detection in formalin fixed paraffin embedded human tissue by immunohistochemistry. Eur. J. Histochem., 59.","DOI":"10.4081\/ejh.2015.2481"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"59058","DOI":"10.18632\/oncotarget.19939","article-title":"Beclin 1 and LC3 as predictive biomarkers for metastatic colorectal carcinoma","volume":"8","author":"Zhao","year":"2017","journal-title":"Oncotarget"}],"container-title":["Cancers"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6694\/11\/3\/389\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T12:39:08Z","timestamp":1760186348000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6694\/11\/3\/389"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,3,19]]},"references-count":35,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2019,3]]}},"alternative-id":["cancers11030389"],"URL":"https:\/\/doi.org\/10.3390\/cancers11030389","relation":{},"ISSN":["2072-6694"],"issn-type":[{"value":"2072-6694","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,3,19]]}}}