High Beclin 1 expression defines a poor prognosis in endometrial adenocarcinomas

doi:10.1016/j.ygyno.2011.06.023

Gynecologic Oncology

Volume 123, Issue 1, October 2011, Pages 147-151

https://doi.org/10.1016/j.ygyno.2011.06.023 Get rights and content

Abstract

Objective

To investigate the prognostic role of Beclin 1 in endometrial adenocarcinomas of the endometrioid cell type. Beclin 1 is a known tumor suppressor gene, but its function may be altered under conditions of an accelerated autophagic activity, which provides additional energy to proliferating cells by recycling defective organelles and long-lived cytoplasmic proteins.

Materials and methods

One hundred and fifty-five endometrioid adenocarcinomas were investigated for their autophagic activity using the monoclonal antibody Beclin 1 and an automated immunohistochemical technique. The extent of Beclin 1 expression was evaluated on a three-tier scale as follows: low (< 10% positive tumor cells), intermediate (between 10% and 50% positive tumor cells), and high (> 50% positive tumor cells). The results were correlated with the degree of tumor differentiation, the depth of myometrial invasion and the overall 5-year survival. In addition, the endometrial tumors were immunostained with the hypoxia inducible factor 1α (HIF1α) and their expression was related to Beclin 1.

Results

A high Beclin 1 reactivity occurred in 18.1% of endometrial adenocarcinomas studied and was associated with high tumor grade, high myometrial invasion and a poor 5-year survival. It was also correlated positively with HIF1α. Of the remaining adenocarcinomas 29.7% were of intermediate Beclin 1 reactivity and 52.2% of low, but correlations with prognostic factors were insignificant.

Conclusion

An increased Beclin 1 expression is connected with the most aggressive endometrioid adenocarcinomas, probably as a result of its strong association with tumor hypoxia.

Highlights

► Beclin 1, a marker of autophagic activity, was highly expressed in 18% of endometrioid adenocarcinomas and down-regulated in 52%. ► A high Beclin 1 expression was associated with high tumor grade, deep myometrial invasion and a poor 5-year survival. ► HIF1α and Beclin 1 proteins were significantly co-expressed, suggesting a link between beclin 1 expression and intratumoral hypoxia.

Introduction

Autophagy is an intracellular homeostatic mechanism, most important for the recycling of long-lived cytoplasmic proteins and defective organelles [1]. The formation of autophagosomes, that is double membrane vacuoles engulfing cytoplasmic components, is the first step in the autophagic process. These are, subsequently, fused with lysosomes (autolysosomes) and the sequestered material is degraded by lysosomal enzymes. Thus, the intracellular accumulation of waste constituents is prevented and, at the same time, complex molecules are converted to simpler for cell metabolism.

Given that various stress conditions, such as hypoxia and high acidity, accentuate the autophagic machinery, it is possible that, under these circumstances, autophagy may assume a dominant role in cancer cell survival and growth [2], [3].

Beclin 1 (BECN1), a mammalian orthologue of the yeast Apg6/Vps30 gene [4], functions as a scaffold for the formation of the PI3K complex, which is one of the first components recruited during the development of autophagosomes. It has, therefore, an important role in autophagy. Furthermore, Beclin 1 is present in intra-cytoplasmic organelles, including the trans-Golgi network, the endoplasmic reticulum, the mitochondria and the perinuclear membrane [4], [5].

In a previous study [6], we have shown that the LC3A protein, an essential component of the autophagic vacuoles, is highly expressed in a proportion of endometrial carcinomas and that a specific pattern reflecting excessive autophagic activity, namely the ‘stone-like’ structures (SLS), are linked with poor prognosis. In this study, we investigate the expression of Beclin 1 in a series of 155 endometrial adenocarcinomas of the endometrioid cell type in order to examine its clinico-pathological and prognostic role in malignant endometrial disease.

Section snippets

Materials and methods

The material comprised 155 endometrial adenocarcinomas of the endometrioid cell type, and 20 normally cycling endometria. The tissues, which were drawn from the files of the Department of Pathology, Democritus University of Thrace, Alexandroupolis, Greece, had been routinely fixed in 10% formol-saline.

All cancer patients in the series had been treated with total abdominal hysterectomy and bilateral salpingo-oophorectomy. Their age ranged from 47 to 80 years (median 63). Patients with myometrial

Beclin 1 reactivity in endometrial tumor cells

Endometrial tumor cells expressed high Beclin 1 reactivity in 18.1% (28/155) of the cases studied (Fig. 1b); reactivity was intermediate in 29.7% (46/155), and low in 52.2% (81/155).

Beclin 1 reactivity in the normal endometrium

All normally cycling endometria, whether proliferating or secretory, demonstrated a high Beclin 1 reactivity in glandular epithelial cells, while stromal cells were unreactive.

Association with histological variables and HIF-1α

The association of Beclin 1 with the various histopathological parameters studied is shown in Table 1. High Beclin 1 expression was

Discussion

Beclin 1 is an essential protein for the initiation of autophagosome formation [4]. It resides in the trans-Golgi network, the endoplasmic reticulum and the mitochondria [4], [5], and a cytoplasmic reactivity, after immunohistochemical stain, was not, therefore, unexpected in both normal and malignant endometrial tissues.

Indeed, Beclin 1 was expressed strongly in the normally cycling endometrium, compared to stromal and myometrial cells, suggesting a role of the autophagic machinery in the

Conflict of interest statement

The authors declare that there are no conflicts of interest.

Acknowledgments

The authors thank Professor K.C. Catter, University of Oxford, Oxford, UK for kindly providing the monoclonal antibody HIF1α. They also thank Mrs. Kyriaki Devetzi for excellent technical assistance.

References (36)

R.J. Lotocki et al.
Stage I endometrial adenocarcinoma: treatment results in 835 patients
Am J Obstet Gynecol
(1983)
R.W. Noyes et al.
Dating the endometrial biopsy
Fertil Steril
(1950)
M.I. Koukourakis et al.
Hypoxia-inducible factor (HIF1A and HIF2A), angiogenesis, and chemoradiotherapy outcome of squamous cell head-and-neck cancer
Int J Radiat Oncol Biol Phys
(2002)
C.H. Yeh et al.
Hypoxic preconditioning reinforces HIF-alpha-dependent HSP70 signaling to reduce ischemic renal failure-induced renal tubular apoptosis and autophagy
Life Sci
(2010)
D.J. Klionsky et al.
Autophagy as a regulated pathway of cellular degradation
Science
(2000)
M.T. Rosenfeldt et al.
The multiple roles of autophagy in cancer
Carcinogenesis
(2011)
E. White et al.
The double-edged sword of autophagy modulation in cancer
Clin Cancer Res
(2009)
Y. Cao et al.
Physiological functions of Atg6/Beclin 1: a unique autophagy-related protein
Cell Res
(2007)
A. Kihara et al.
Beclin-phosphatidylinositol 3-kinase complex functions at the trans-Golgi network
EMBO Rep
(2001)
E. Sivridis et al.
Autophagy in endometrial carcinomas and prognostic relevance of ‘stone-like’ structures (SLS): what is destined for the atypical endometrial hyperplasia?
Autophagy
(2011)

S.G. Silverberg et al.

Tumors of the Uterine Corpus and Gestational Trophoblastic Disease

(1992)

W.T. Creasman

Announcement. FIGO stages—1988 revision

Gynecol Oncol

(1989)

R.J. Zaino et al.

The utility of the revised International Federation of Gynecology and Obstetrics histologic grading of endometrial adenocarcinoma using a defined nuclear grading system. A Gynecologic Oncology Group study

Cancer

(1995)

Carcinoma of the endometrium

ACOG Technical Bulletin Number 162–December 1991

Int J Gynaecol Obstet

(1993)

S. Biesterfeld et al.

Analysis of the reliability of manual and automated immunohistochemical staining procedures. A pilot study

Anal Quant Cytol Histol

(2003 Apr)

D.Y. Lu et al.

Automated in situ hybridization and immunohistochemistry for cytomegalovirus detection in paraffin-embedded tissue sections

Appl Immunohistochem Mol Morphol

(2009)

S. Arbogast et al.

Automated ERCC1 immunohistochemistry in non-small cell lung cancer: comparison of anti-ERCC1 antibodies 8F1, D-10, and FL-297

Appl Immunohistochem Mol Morphol

(2011)

E. Sivridis et al.

Association of hypoxia-inducible factors 1alpha and 2alpha with activated angiogenic pathways and prognosis in patients with endometrial carcinoma

Cancer

(2002)

Cited by (63)

Atg9A-mediated mitophagy is required for decidual differentiation of endometrial stromal cells
2022, Reproductive Biology
Citation Excerpt :
However, ATG9A is the only integral autophagy membrane protein essential for autophagosome formation [16]. Silencing of Beclin-1 (BECN1), considered a marker of poor prognosis for endometrial adenocarcinomas, could reduce autophagic flux in the endometrium [17,18]. The purpose of this study was to identify molecular mechanisms underlying differentiation of endometrial stromal fibroblasts into specialized secretory decidual cells.
Endometrial decidualization is the foundation of a healthy pregnancy. Mitochondrial dysfunction is an independent cause of disease for energy-intensive organs. Mitochondrial homeostasis plays a key role in the differentiation processes of many cell types. We showed increased activation of mitophagy (mitochondrial autophagy) in the decidua compared with proliferative or secretory endometrium. To better comprehend the mechanisms underlying healthy conception, understanding the mechanism of endometrial stromal cell decidualization is of great importance. Here, we artificially induced decidualization of a human endometrial stromal cell line (T HESCs) and characterized subsequent activation of mitophagy using immunofluorescence assay, electron microscopy and Western blot assay. Knockdown of autophagy-related 9A (ATG9A) led to an obvious reduction of mitophagy and deficiencies in decidualization. Our findings demonstrate a key role for proper mitochondrial dynamics in decidual differentiation and identify ATG9A-mediated mitophagy as a novel therapeutic target for repeated implantation failure or recurrent abortion.
Beclin1-mediated interplay between autophagy and apoptosis: New understanding
2022, International Journal of Biological Macromolecules
Citation Excerpt :
In these malignant cells, the Beclin1 is associated with another protein 14–3-3ζ under hypoxia (2% oxygen) condition, leading to cause enhanced autophagy due to chemoresistance, suggesting the tumor-promoting role of Beclin1 in primary liver cancer [68]. Similar effects of Beclin1 have been shown in other forms of cancer, such as ovarian carcinogenesis [69] and endometrial adenocarcinomas [70]. Furthermore, in non-small cell lung cancer (NSCLC), the overexpression of Beclin1 has been correlated with enhanced migration of NSCLC cells, and the Beclin1 upon knockdown results in reduced migratory ability.
The definition for autophagy holds a ‘single’ meaning as a conserved cellular process that constitutes a recycling pathway for damaged organelles and long-lived proteins to maintain nutrient homeostasis and mediate quality control within the cell. But this process of autophagy may behave ambiguously depending on the physiological stress as the stress progresses in the cellular microenvironment; the ‘single’ meaning of the autophagy changes from the ‘cytoplasmic turnover process’ to ‘tumor suppressive’ and a farther extent, ‘tumor promoter’ process. In a tumorigenic state, the chemotherapy-mediated resistance and intolerance due to upregulated autophagy in cancer cells have become a significant concern. This concern has provided insight to the scientific community to enter into the arena of cross-talk between autophagy and apoptosis. Recent findings and ongoing research have provided insights on some of the key regulators of this cross-talk; one of them is Beclin1 and their involvement in the physiological and the pathophysiological processes; however, reconciliation of these two forms of death remains an arena to be explored extensively. This review sheds light on the interplay between autophagy and apoptosis, emphasizing one of the key players, Beclin1, and its importance in health and diseases.
A panel of eight autophagy-related long non-coding RNAs is a good predictive parameter for clear cell renal cell carcinoma
2021, Genomics
Clear-cell renal cell carcinoma (ccRCC) carries a variable prognosis. Prognostic biomarkers can stratify patients according to risk, and can provide crucial information for clinical decision-making. We screened for an autophagy-related long non-coding lncRNA (lncRNA) signature to improve postoperative risk stratification in The Cancer Genome Atlas (TCGA) database. We confirmed this model in ICGC and SYSU cohorts as a significant and independent prognostic signature. Western blotting, autophagic-flux assay and transmission electron microscopy were used to verify that regulation of expression of 8 lncRNAs related to autophagy affected changes in autophagic flow in vitro. Our data suggest that 8-lncRNA signature related to autophagy is a promising prognostic tool in predicting the survival of patients with ccRCC. Combination of this signature with clinical and pathologic parameters could aid accurate risk assessment to guide clinical management, and this 8-lncRNAs signature related to autophagy may serve as a therapeutic target.
Crosstalks between Yin-Yang 1 (YY1) and autophagy in cancer
2020, Autophagy in Immune Response: Impact on Cancer Immunotherapy
The transcription factor Yin Yang 1 (YY1) is overexpressed in the majority of human cancers and is closely linked in the regulation of the various phenotypic properties of cancer. YY1 regulates cell viability, cell proliferation, migration, invasion, EMT, metastasis, and the resistance to chemo-immuno-therapeutic cytotoxic drugs. Its overexpression has also been associated with poor prognosis. Autophagy is a cellular process that cancer cells regulate for their own survival under stress conditions. Hence, both YY1 and autophagy share common properties in cancer and, therefore, it was hypothesized that YY1 may regulate autophagy in cancer via several crosstalk signaling pathways. The analyses identified, indeed, the presence of multiple crosstalks that primarily consisted of the followings: (1) the YY1/p53 autophagy axis, (2) the YY1/Atg genes autophagy axis, (3) the YY1/LC3 autophagy axis, (4) the YY1/miR 30a autophagy axis, (5) the YY1/miR 372 autophagy axis, and (6) the dysregulated NF-kB/SNAIL/YY1/RKIP/PTEN autophagy axis. Clearly, these various crosstalks supported the proposed hypothesis and demonstrated an additional activity for YY1 in the regulation of autophagy in cancer. The implication of these findings is that targeting either the crosstalk pathways and/or directly YY1 may be detrimental to the tumor cell survival, growth, invasion, metastasis, resistance and, subsequently, leading to tumor regression.
Inhibition of autophagy by chloroquine makes chemotherapy in nasopharyngeal carcinoma more efficient
2019, Auris Nasus Larynx
A combination of platinum-based chemotherapy and radiotherapy is the standard treatment for nasopharyngeal carcinoma (NPC). However, the efficacy of chemotherapy has reached a plateau. Many autophagy studies suggest that autophagy can either promote or suppress to cancer progression. Thus, a role of autophagy in the acquisition of chemoradioresistance has recently been a notable event. Therefore, we examined the relationship between autophagy and chemotherapy in NPC.
The expression of Beclin 1 and microtubule-associated protein light chain 3 (LC3), a marker of autophagy, was determined by immunohistochemistry in the biopsy samples of patients with NPC before and after the first course of chemotherapy. Additionally, to investigate in the effect of autophagy suppression in chemotherapy, NPC cell line C666-1 cells were treated with cisplatin and/or chloroquine, an inhibitor of autophagy.
The expression of Beclin 1 increased after chemotherapy in all patients. In NPC cell line C666-1, compared to cisplatin alone, combination therapy (cisplatin and chloroquine) reduced cell viability, and promoted cell apoptosis.
These results suggest that autophagy, represented by Beclin 1, is upregulated after chemotherapy in both in vitro and in vivo NPC studies. Inhibition of autophagy could therefore be new strategy for NPC treatment.
Phytochemical Modulation of Apoptosis and Autophagy: Strategies to Overcome Chemoresistance in Leukemic Stem Cells in the Bone Marrow Microenvironment
2017, International Review of Neurobiology
Citation Excerpt :
Initially, autophagy likely has a preventive effect against cancer, but as the cancer develops, the cancer cells could utilize autophagy for their own cytoprotection, probably due to the role of this process in improving nutrient and/or oxygen supply to rapidly dividing cancer cell populations (Janku, McConkey, Hong, & Kurzrock, 2011). Indeed, the aberrant expression of BECLIN1 has been reported in many types of cancer including hepatocellular, colorecetal, nasopharyngeal, endometrial, and gastric (Giatromanolaki et al., 2011; Koukourakis et al., 2010; Shi, Ding, Zhou, Qiu, & Fan, 2009; Wan et al., 2010; Xia, Wang, Zhao, & Song, 2013) and has been shown to correlate with poor prognosis. Recently, autophagy has been shown to play a role in the chemoresistance of CML stem cells to TKIs.
Advances in scientific research and targeted treatment regimes have improved survival rates for many cancers over the past few decades. However, for some types of leukemia, including acute lymphoblastic and acute myeloid leukemia, mortality rates have continued to rise, with chemoresistance in leukemic stem cells (LSCs) being a major contributing factor. Most cancer drug therapies act by inducing apoptosis in dividing cells but are ineffective in targeting quiescent LSCs. Niches in the bone marrow, known as leukemic niches, behave as “sanctuaries” where LSCs acquire drug resistance. This review explores the role of the bone marrow environment in the maintenance of LSCs and its contribution to chemoresistance and considers current research on the potential use of phytochemicals to overcome chemoresistance through the modulation of signaling pathways involved in the survival and death of leukemic clonal cells and/or leukemic stem cells. Phytochemicals from traditional Chinese medicine, namely baicalein, chrysin, wogonin (constituents of Scutellaria baicalensis; huáng qín; 黄芩), curcumin (a constituent of Curcuma longa, jiāng huáng, 姜黄), and resveratrol (a constituent of Polygonum cuspidatum; hŭ zhàng, 虎杖) have been shown to induce apoptosis in leukemic cell lines, with curcumin and resveratrol also causing cell death via the induction of autophagy (a nonapoptotic pathway). In order to be effective in eliminating LSCs, it is important to target signaling pathways (such as Wnt/β-catenin, Notch, and Hedgehog). Resveratrol has been reported to induce apoptosis in leukemic cells through the inhibition of the Notch and Sonic hedgehog signaling pathways, therefore showing potential to affect LSCs. While these findings are of interest, there is a lack of reported research on the modulatory effect of phytochemicals on the autophagic cell death pathway in leukemia, and on the signaling pathways involved in the maintenance of LSCs, highlighting the need for further work in these areas.

View all citing articles on Scopus

View full text

High Beclin 1 expression defines a poor prognosis in endometrial adenocarcinomas

Abstract

Objective

Materials and methods

Results

Conclusion

Highlights

Introduction

Section snippets

Materials and methods

Beclin 1 reactivity in endometrial tumor cells

Beclin 1 reactivity in the normal endometrium

Association with histological variables and HIF-1α

Discussion

Conflict of interest statement

Acknowledgments

Am J Obstet Gynecol

Fertil Steril

Int J Radiat Oncol Biol Phys

Life Sci

Autophagy as a regulated pathway of cellular degradation

Science

The multiple roles of autophagy in cancer

Carcinogenesis

The double-edged sword of autophagy modulation in cancer

Clin Cancer Res

Physiological functions of Atg6/Beclin 1: a unique autophagy-related protein

Cell Res

Beclin-phosphatidylinositol 3-kinase complex functions at the trans-Golgi network

EMBO Rep

Autophagy in endometrial carcinomas and prognostic relevance of ‘stone-like’ structures (SLS): what is destined for the atypical endometrial hyperplasia?

Autophagy

Tumors of the Uterine Corpus and Gestational Trophoblastic Disease

Announcement. FIGO stages—1988 revision

Gynecol Oncol

The utility of the revised International Federation of Gynecology and Obstetrics histologic grading of endometrial adenocarcinoma using a defined nuclear grading system. A Gynecologic Oncology Group study

Cancer

ACOG Technical Bulletin Number 162–December 1991

Int J Gynaecol Obstet

Analysis of the reliability of manual and automated immunohistochemical staining procedures. A pilot study

Anal Quant Cytol Histol

Automated in situ hybridization and immunohistochemistry for cytomegalovirus detection in paraffin-embedded tissue sections

Appl Immunohistochem Mol Morphol

Automated ERCC1 immunohistochemistry in non-small cell lung cancer: comparison of anti-ERCC1 antibodies 8F1, D-10, and FL-297

Appl Immunohistochem Mol Morphol

Association of hypoxia-inducible factors 1alpha and 2alpha with activated angiogenic pathways and prognosis in patients with endometrial carcinoma

Cancer