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PYGO2 regulates IL10 and plays immunosuppressive role through ESCC progression

BMC Molecular and Cell Biology volume 26, Article number: 14 (2025) Cite this article

Abstract

Background

Esophageal squamous cell carcinoma (ESCC), one of the most aggressive carcinomas of the gastrointestinal tract, is the sixth most common cause of cancer-related death. Wnt pathway plays a pivotal role in cell proliferation and differentiation. PYGO2 and IL10 are involved in this pathway. Our aim in this study was to examine the correlation between PYGO2 and IL10 expression in ESCC patients and cell lines.

Methods

Relative-comparative real time-PCR (RT-qPCR) was used to evaluate the PYGO2 and IL10 mRNA expression profile in 58 non-treated ESCC compared to their margin normal tissues. Expression of PYGO2 was induced in KYSE-30 and YM1 ESCC lines and IL10 expression was analyzed.

Results

The results revealed the significant overexpression of PYGO2 and IL10 mRNA in 31.0% and 51.7% of ESCCs, respectively. The PYGO2 and IL10 overexpression was significantly correlated to each other (p = 0.007). Concomitant overexpression of the genes was significantly associated to grade of tumor differentiation (p < 0.01), and tumor depth of invasion (p < 0.05). Induced expression of PYGO2 caused a meaningful change in IL10 expression in ESCC cells.

Conclusion

PYGO2 may regulate IL10 through Wnt/β-catenin signaling pathway, suggesting a possible oncogenic role for PYGO2/IL10 axis in ESCC tumorigenesis. Considering the involvement of IL10 as an anti-inflammatory cytokine and PYGO2 role in elevated tumor invasion and metastasis, possible functional interaction between these factors may promote a process which induces invasion and malignant phenotype in ESCC.

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Introduction

Esophageal cancer (EC) is the 8th most prevalent malignancy and 6th leading cause of cancer-related death worldwide. It can be histopathologically categorized to two common subtypes including esophageal squamous cell carcinoma (ESCC) and esophageal adenocarcinoma (EAC). ESCC as the predominant subtype of EC, accounts for nearly 90% of EC particularly in Iran, North China, and Africa. Due to late diagnosis, remote metastasis, and local invasion, ESCC has a quite poor 5-year survival rate (15–25%), and patients have to relinquish the principal curative option of surgical resection [1, 2].

Deregulation of signaling pathways, such as canonical Wnt/β-catenin, is associated with the progression and development of various malignancies. Binding of specific Wnt ligands to membrane receptors/co-receptors induces inactivation of an AXIN-APC-GSK3β destruction complex which results in stabilization and accumulation of β-catenin in the cytoplasm. Consequently, β-catenin is translocated into the nucleus and binds to TCF/LEF transcription factors to activate transcription of Wnt target genes [3]. Pygopus2 (PYGO2), which contains a highly conserved C-terminal plant homeo domain (PHD), functions as a significant coactivator of the Wnt/β-catenin transcriptional complex (LEF/TCF). PYGO2 directly bind to histone H3 trimethylated at lysine 4 (H3K4me3), an epigenetic mark linked to transcriptional activation, through its PHD and activates β-catenin-dependent transcriptional regulation [4]. Due to PYGO2 substantial regulatory roles in the cell, its deregulation may lead to tumorigenesis. Pivotal role of PYGO2 has revealed in multiple cancers including ESCC, ovarian, breast, cervical, colon, lung, and liver malignancies.

According to contribution of chronic inflammation in EC susceptibility, several involved genes in inflammatory pathways may play role in EC development. Interleukin (IL)-10, an immunosuppressive T helper 2 type cytokine, is secreted by various types of cells including B cells, dendritic cells, monocytes/macrophages, diverse T regulatory cell subsets, CD8+ and CD4+ T cells and natural killer (NK) cells. IL10 plays diverse roles in the regulation of immune activities such as inhibition of T helper 1 type cytokine production and T lymphocyte proliferation, blunting of cytotoxic responses, and impairment of antigen presentation cells. The regulation of IL10 expression appears to be intricate, so that it can function as either a pro-tumorigenic or an antitumor agent [5]. However, upregulation of IL10 expression has verified in a number of hematopoietic and solid tumors including ESCC, colon, breast, and lung cancers [6, 7]. The Wnt/β-catenin signaling pathway applies both proinflammatory and anti-inflammatory functions. There was a remarkable correlation between enhancing the level of Wnt pathway ligand/receptor (Wnt3a/FZD9, respectively) and increased level of IL10 in enteric nervous system [8]. IL10 has likewise been demonstrated as a β-catenin target gene in colon cancer [9]. These findings support the involvement of IL10 in Wnt/β-catenin pathway active state.

Considering the aberrant activation of Wnt/β-catenin signaling pathway in progression and metastasis of multiple cancers, as well as involvement of PYGO2 and IL10 gene expression in ESCC development, we aimed in the present study to evaluate the probable correlation between PYGO2 and IL10 expression in ESCC patients and cell lines.

Materials and methods

In silico study

The GeneMANIA web-based database was employed to predict potential targets of PYGO2-IL10 through key genes of the Wnt/β-catenin signaling pathways (https://genemania.org/). Additionally, the Phenolyzer website was utilized to predict interactions between PYGO2-IL10, as well as key genes in the Wnt/β-catenin pathway, within the context of ESCC (https://phenolyzer.wglab.org/).

Cell lines and culture condition

KYSE-30 and YM1 ESCC lines were obtained from Pasteur Institute of Iran (Tehran, Iran), and cultured in RPMI-1640 and DMEM culture mediums (Bio-Idea, Iran) supplemented with 10% fetal bovine serum (FBS solution, BI, USA), and 100 µg/ml penicillin/streptomycin (Gipco, USA), and incubated in %90 humidity incubator containing %5 CO2 at 37 °C.

Transfection

The Human PYGO2 ORF expression plasmid, C-HA tag (Sino Biological Inc. Catalog Number: HG20275-CY) was applied to enforce PYGO2 expression in the cells. Transfection was performed using Lipofectamin 2000 transfection reagent (Roche, Basel, Switzerland). Nearly 600,000 cells were seeded per six-well plate and transfected with a total of 2 µg plasmid. The transfection efficiency was checked [10]. After 48 h cells were treated with trypsin and EDTA and subjected to RNA extraction.

ESCC patients and tissue samples

The fresh tumoral and para-tumor normal tissues were collected from 58 ESCC patients who underwent surgical removal of tumor tissue in Omid Hospital of Mashhad University of Medical Sciences (MUMS). These patients had not received radiotherapy or chemotherapy before the surgery. All patients provided the informed consent and the study was approved by the Ethics Committee of MUMS (No.88098). According to the latest Union International Cancer TNM classification guidelines, the histopathological features of the samples including tumor size, nodal involvement, and metastasis (TNM) staging were determined, then samples were confirmed to be tumor or normal histologically.

RNA extraction, cDNA synthesis and quantitative RT-PCR

RNA was isolated from all tissues, using TriPure RNA extraction reagent (Roche, Nutley, NJ). The RNA integrity in all samples were verified by agarose gel electrophoresis. The cDNA was synthesized from RNA with oligo (dT)18 and random hexamer primers concurrently. Relative comparative real-time PCR for PYGO2 and IL10 genes were performed with peculiar primer sets (Table 1) using SYBR Green method, containing ROX as a reference dye in a Stratagene Mx3000P real-time thermocycler detection system (Stratagene, La Jolla, CA, USA). The thermal cycling program included an initial denaturation step of 10 min at 95 °C, followed by 40 cycles of 15 s at 95 °C, 30 s at 62 °C, and 45 s at 72 °C. All samples were tested in triplicate. For the normalization of RNA input, Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was employed as an endogenous control, which displays the lowest mutability in expression in esophageal tissues [11]. The ΔΔCt method was employed to evaluate fold change of gene expression, as described before.

Table 1 Primer sequences used for comparative real-time RT-PCR
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Statistical analysis

The statistical analyses were performed using SPSS 19.0 statistical package (SPSS, Chicago, IL, USA). The χ2 or Fisher’s exact test, independent-sample t test and ANOVA were utilized to analyze associations between gene expression and diverse histopathological factors. The correlation between PYGO2 and IL10 expression was analyzed using Pearson’s correlation. A p-value of < 0.05 was defined as statistically significant.

Results

PYGO2-IL10 pro-pro interaction with key genes of Wnt signaling pathway

In this study, we performed comprehensive functional analyses to investigate the interaction between PYGO2-IL10 and key genes involved in the Wnt signaling pathway. To assess the interactive relationships, we utilized open-source expression analysis tools, including GeneMania25 and phenolyzer. Figure 1a illustrates the results of the interactive analysis, focusing on PYGO2-IL10, and the expression of Wnt signaling pathway genes. GeneMANIA analysis revealed significant protein-protein interactions between PYGO2 and a variety of Wnt signaling proteins, including CREBBP, BCL9, CTNNB1, KMT2D and PYGO1. Notably, pivotal protein-protein interactions were also observed between IL10 and SOX2, PTK7, IL26, IL19, IL20 and IL10RA, as depicted in Fig. 1.

Fig. 1
figure 1

(a) The predicted protein-protein interaction network generated using GeneMANIA, illustrating the predicted relationships between PYGO2-IL10 through Wnt signaling pathway genes. (b) The protein-protein interaction network for PYGO2. (c) The protein-protein interaction network for IL10. (d) The predicted protein-protein interaction network in the context of ESCC, based on predictions from Phenolyzer

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Furthermore, phenolyzer analyses were conducted to explore the contextual associations of PYGO2-IL10 with various Wnt signaling pathway key genes in the context of ESCC. The results obtained from phenolyzer analyses (Fig. 1d) highlighted a multitude of significant cellular associations linking PYGO2-IL10 with these genes. These findings provide valuable insights into the functional interactions of PYGO2-IL10 with key genes within the Wnt signaling pathway.

ESCC patients and tissue samples

Primary frozen tumor tissues and their margin non-tumorous esophagus were obtained from 58 newly diagnosed ESCC patients prior to receive radio- or chemo-therapy. Thus, the histopathological characteristics of ESCC tissues were not affected by subsequent therapeutic interventions. The male-to-female ratio of enrolled patients in this study was 1.2 (32:26). The mean age ± standard deviation (SD) of patients was 61.85 ± 12.25 (age range 30–83 years). The size of tumor samples ranged from 0.5 to 12 cm (mean ± SD: 4.01 ± 1.93), resected from middle or lower parts of the esophagus. The clinicopathological features of the patients are listed in Table 2.

Table 2 Correlations between PYGO2 and IL10 gene expression and clinicopathological characteristics of ESCC patients
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Upregulation of PYGO2 and IL10 in ESCC

Relative comparative real-time PCR (qRT-PCR) was used to analyze PYGO2 and IL10 expression in 58 ESCC patients. Both genes were significantly overexpressed at mRNA level in 18 of 58 (31.0%) and 30 of 58 (51.7%) ESCC tumors compared to paired adjacent non-tumor tissues, respectively. The mean (± SD) of PYGO2 mRNA expression fold changes in ESCC tissues was 1.02 (± 2.18), with minimum and maximum fold changes of -3.31 and 7.00, respectively. The mean (± SD) of IL10 fold changes in ESCC tissues was 1.81 (± 2.39). Its fold changes was ranged from − 4.50 to 6.03 fold, in ESCC samples. Figure 2 visualizes the profile of PYGO2 and IL10 gene expression in ESCC patients as a scatter plot.

Fig. 2
figure 2

Descriptive analysis of PYGO2 and IL10 gene expression pattern in ESCC patients as scatter plot. The Y-axis represents the log 2 fold change of gene expression, and the X-axis indicates the number of patients. Relative mRNA expression of more than two-fold in tumor tissue is considered as overexpression; less than minus two-fold as underexpression and the range in between is defined as normal or no change

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Association between PYGO2 and IL10 gene expression

The PYGO2 and IL10 expression were significantly correlated to each other in ESCC samples (p = 0.007, correlation coefficient = 0.385). Regression plot depicting this correlation is presented in Fig. 3. Intriguingly, gene expression data from ESCC samples indicated the overexpression of both genes in 24.13% (14 of 58) of patients, whereas 34.48% (20 of 58) of specimens had high mRNA level of at least one gene. Neither PYGO2 nor IL10 was overexpressed in 41.37% (24 of 58) of tumors (Table 3).

Fig. 3
figure 3

Regression plot representing correlation between PYGO2 and IL10 mRNA expression (p = 0.007). X and Y axises show log 2 fold change of gene expression

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Table 3 Association between PYGO2 and IL10 expression in ESCC patients
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Association of the genes with surgical stage

The overexpression of PYGO2 and IL10 was associated to each other in higher-stage of the disease (stage III; p < 0.01). Indeed, 7 of 18 tumors (38.88%) that overexpressed PYGO2 and 9 of 30 tumors (30.0%) that overexpressed IL10 showed stage III of tumor progression. While 36% (21 of 58) of tumors with advanced stage of disease displayed significant co-overexpression of PYGO2 and IL10 (p < 0.001), other cases with lower tumor stages (I and II) did not manifest co-overexpression of the genes (p = 0.189).

Association of the genes with lymph node metastasis

Interestingly, we found significant correlation between expression of the genes in metastasized tumor cell into the lymph nodes (p = 0.001). Moreover, PYGO2 overexpression was significantly associated with tumor metastasis to lymph nodes (p = 0.017). Among 25 patients with node metastasis, 7 patients (28%) had PYGO2 overexpression. However, in 66.66% of patients (22 of 33) without any metastasis to lymph nodes, PYGO2 was not overexpressed. There was no significant association between IL10 overexpression and lymph node metastasis.

Association of the genes with depth of tumor invasion

The expression pattern of both genes was significantly associated to each other in ESCC patients with invaded tumors to the adventitia (T3; p = 0.031). Concomitant overexpression of the genes was significantly correlated with depth of tumor invasion (p = 0.032). Overexpression of IL10 was also significantly correlated with depth of tumor invasion (p = 0.020). Indeed, 25 of 47 (53.19%) tumors with invasion to adventitia (T3), showed IL10 overexpression.

Association of the genes with grade of tumor differentiation and location

We found a significant association between PYGO2 and IL10 expression in moderately differentiated tumor samples (p = 0.001), but not in patients with well and poorly differentiated tumors (p = 0.102). Co-overexpression of PYGO2 and IL10 was displayed in 9 of 35 (25.7%) samples with moderately differentiated tumors whereas in patients with well and poorly differentiated tumors, this percentage was 1 of 12 (8.3%) and 4 of 11 (36.4%), respectively. PYGO2 and IL10 expression were lowest in well-differentiated tumors compared to moderately differentiated tumors. Besides, tumor samples from the middle region of esophagus expressed IL10 at higher levels than lower region tumors. There were no significant correlations between PYGO2 and IL10 expression and other clinicopathological variables.

Forced expression of PYGO2 changed IL10 expression

After the transfection of KYSE-30 and YM1 cells, a functional study was performed to evaluate the expression of PYGO2 and IL10. PYGO2 was overexpressed in KYSE-30 and YM1 cells compared to controls, with log 2 fold changes nearly 6.5 and 4.8, respectively. This level of PYGO2 overexpression caused a significant increase in IL10 gene expression nearly 4 and 2.5 in KYSE-30 and YM1 cells, respectively.

Discussion

The Wnt/β-catenin signaling pathway, as an evolutionarily-conserved pathway, employs both proinflammatory and anti-inflammatory functions [12]. Its activation is associated with deficiency of an immune infiltrate in cancer microenvironment [13]. Deregulation of this pathway was detected in liver, osteosarcoma, breast, colorectal, gastric, cervical, and bone cancers [14]. Consist with these findings, several reports corroborate activation of this pathway in ESCC.

Here we manifested that mRNA expression of PYGO2 and IL10, as two important genes involved in Wnt signaling pathway, was upregulated in ESCC and intriguingly, there was a significant correlation between these genes which probably occur through Wnt/β-catenin signaling pathway. Having performed functional study, we demonstrated that enforced expression of PYGO2 changed IL10 expression in ESCC cells. Furthermore, we found that concomitant expression of the genes was strongly correlated with clinical features of ESCC patients and different indices of poor prognosis which may introduce their concomitant expression as a new regulatory axis in ESCC.

PYGO2 is an essential transcriptional coactivator of the Wnt/β-catenin signaling pathway which promotes β-catenin-LEF/TCF transcriptional activation through adapter protein BCL9 [15]. It contributes in expression of highly transcribed RNAs during cell-cycle progression and DNA replication, growth and expansion of cancer cells, as well as histone code interpretation through β-catenin– histone methyltransferase/histone acetyltransferase (HMT/HAT) association [16]. Furthermore, it is applied in a MYC-dependent transcriptional program as an epigenetic accessory protein to regulate cell division [17]. Up-regulated expression of PYGO2 has been illustrated in diverse malignant tumors including ovarian, colon, liver, and cervical cancers. In addition, its oncogenic role is confirmed in hepatic carcinoma, prostate adenocarcinoma, lung and breast cancers as well as renal cell carcinoma and glioma [18,19,20,21]. In line with these reports, PYGO2 overexpression has been reported in ESCC in association with the grade of tumor cell differentiation [22]. Intriguingly, we also observed high level expression of PYGO2 in 31.0% of ESCC samples. It has been revealed that PYGO2 overexpression promotes lymph node metastasis in lung and prostate cancers [23]. Here we found that 7 of 18 PYGO2 overexpressed tumors, were metastasized to the lymph nodes, probably reflecting the importance of PYGO2 in metastasis process. Likewise, PYGO2 activates Wnt target genes such as AXIN1, cyclin D1, and IL10. Surprisingly, we found a significant correlation between PYGO2 and IL10 expression. In tumors with high levels of PYGO2, IL10 expression was significantly higher compared to those with normal PYGO2 expression. In addition, induced expression of PYGO2 in ESCC lines increased expression of IL10 significantly. This finding is noteworthy, suggesting the regulatory role of PYGO2 as a transcriptional activator of IL10 expression probably through Wnt/β-catenin signaling pathway (Fig. 4). Since co-overexpression of PYGO2 and IL10 was significantly correlated with different indices of poor prognosis in ESCC, Wnt/β-catenin signaling may be involved in ESCC tumorigenesis through these genes.

Fig. 4
figure 4

Potential correlation between PYGO2 and IL10 expression through the Wnt/β-catenin signaling pathway in ESCC progression. Binding of Wnt ligand to FZD and LRP induces inactivation of the AXIN-APC-GSK3β destruction complex via DVL, leading to β-catenin accumulation and transition to the nucleus. Then, PYGO2 as a coactivator of the transcriptional complex (LEF/TCF/β-catenin), may directly activate transcription of target genes such as IL10. Furthermore, IL10 expression manifests direct correlation with FZD, WNT, LRP expression, β-catenin accumulation, and augmentation of EGFR levels in this pathway

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Cytokines play critical roles in cancer, inducing tumor metastasis and invasion, as well as apoptosis inhibition [24]. Interleukin-10 is a pleiotropic cytokine with broad anti-inflammatory properties through suppression of both dendritic cell and macrophage function. It suppresses the antitumor immune response, so it is indispensable for tumor development [25]. IL10 has widely been examined in various types of human malignancy. Elevated serum concentrations of IL10 is related with poor prognosis and adverse disease stage in gastric, colon, lung, bone sarcoma, hepatocellular carcinoma, melanoma, and pancreatic malignancies. It is interesting that our findings disclosed IL10 overexpression in 51.7% of ESCC samples which is consisting with previous reports in ESCC [26, 27]. Furthermore, we observed 25 of the 30 IL10 overexpressed ESCCs (83.33%) were invaded to the esophagus adventitia presenting T3 depth of tumor invasion. This data corroborates recent reports in melanoma and cervical cancer [28, 29]. According to IL10 function in tumor development, it may be assumed that IL10 induce tumor invasion and progression by suppressing immune responses and stimulating angiogenesis.

IL10 has been illustrated to contribute in divers signaling pathways. It suppresses starvation induced autophagy in HS-derived fibroblasts (HSFs) through cross talk between the IL10/AKT-mTOR and IL10/IL10R-STAT3 pathways [30]. Moreover, Notch signaling can induce IL10 expression through TH1 cells which involves STAT4-dependent process [31]. This gene is closely linked with various Wnt pathway components, wherein multiple Wnt ligands regulate its responses [32]. In addition, fibroblast-derived Wnt16B induces IL10 secretion in dendritic cells. IL10 expression is associated with the levels of EGFR (as a Wnt target gene) and β-catenin accumulation in lung cancer and melanoma, respectively. Similarly, IL10 level is directly correlated with the level of FZD9 and LRP5/6 in Wnt pathway [33,34,35]. Therefore, all these evidences support the involvement of IL10 in Wnt pathway active state.

The concomitant expression of PYGO2 and IL10 in this study was significantly correlated with depth of tumor invasion. Tumor invasion is a predictor of lymph node metastasis in several cancers such as ESCC. Interestingly, of 25 patients with lymph node metastasis, 6 (24.0%) co-overexpressed of both genes, while in 75.8% of patients (25 of 33) without lymph node metastasis, PYGO2 and IL10 were not co-overexpressed. Having consider these results, the correlation between the genes may be envisaged potentially to activate a cascade leading to malignant invasion and metastasis. In the case of tumor stage, the co-overexpression of the genes was significantly correlated with advanced stage of tumor progression (stage III). It was largely expected, since stage III is associated to lymph node involvement. This result confirms the similar findings in NSCLC, where IL10 overexpression was correlated with high degree of stage and lymph node metastases [36]. Considering this fact that ESCC initiation and progression is a multistep process with poor prognosis, the potential correlation between PYGO2 and IL10 may be considered as an efficient prognostic panel of markers in ESCC patients.

Previous studies have separately considered PYGO2 and IL10 as co-transcription factor and target gene in Wnt pathway, respectively. Subsequently, the involvement of deregulated Wnt/β-catenin pathway has been revealed in ESCC carcinogenesis and progression. Based on these reports and our results, it seems likely that PYGO2 may activate IL10 expression through Wnt/β-catenin signaling pathway in ESCC which may augment ESCC carcinogenesis and aggressiveness.

Conclusion

To sum up, our work disclosed the concomitant expression of PYGO2 and IL10 in ESCC patients and regulatory role of PYGO2 in IL10 gene expression in ESCC cells. We indicated an association between these genes probably through Wnt/β-catenin signaling pathway in ESCC. Furthermore, PYGO2 and IL10 co-overexpression was found in significant correlation with tumor poor prognosis including tumor depth of invasion, lymph nodes metastasis, surgical stage, and grade of tumor differentiation. Therefore, it can be introduced as a new regulatory axis for ESCC poor prognosis. To the best of our knowledge, this is the first report found this correlation in ESCC to date. Considering the involvement of IL10 as an anti-inflammatory cytokine and PYGO2 role in elevated tumor invasion and metastasis, possible functional interaction between these factors may promote a process which induces malignant metastasis and invasion in ESCC.

Data availability

All raw data are available on case of reasonable request from corresponding author.

References

  1. Collaborators GBDOC. The global, regional, and National burden of oesophageal cancer and its attributable risk factors in 195 countries and territories, 1990–2017: a systematic analysis for the global burden of disease study 2017. Lancet Gastroenterol Hepatol. 2020;5(6):582–97.

    Article  Google Scholar 

  2. Lagergren J, et al. Oesophageal cancer. Lancet. 2017;390(10110):2383–96.

    Article  PubMed  Google Scholar 

  3. Lustig B, Behrens J. The Wnt signaling pathway and its role in tumor development. J Cancer Res Clin Oncol. 2003;129(4):199–221.

    Article  CAS  PubMed  Google Scholar 

  4. Horsley V. Epigenetics, Wnt signaling, and stem cells: the Pygo2 connection. J Cell Biol. 2009;185(5):761–3.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Mannino MH, et al. The Paradoxical role of IL-10 in immunity and cancer. Cancer Lett. 2015;367(2):103–7.

    Article  CAS  PubMed  Google Scholar 

  6. Galizia G, et al. Prognostic significance of Circulating IL-10 and IL-6 serum levels in colon cancer patients undergoing surgery. Clin Immunol. 2002;102(2):169–78.

    Article  CAS  PubMed  Google Scholar 

  7. Gholamin M, et al. Overexpression and interactions of interleukin-10, transforming growth factor beta, and vascular endothelial growth factor in esophageal squamous cell carcinoma. World J Surg. 2009;33(7):1439–45.

    Article  PubMed  Google Scholar 

  8. Di Liddo R, et al. Anti-inflammatory activity of Wnt signaling in enteric nervous system: in vitro preliminary evidences in rat primary cultures. J Neuroinflammation. 2015;12:23.

    Article  PubMed  PubMed Central  Google Scholar 

  9. Watanabe K, et al. Integrative ChIP-seq/microarray analysis identifies a CTNNB1 target signature enriched in intestinal stem cells and colon cancer. PLoS ONE. 2014;9(3):e92317.

    Article  PubMed  PubMed Central  Google Scholar 

  10. Al AM, Forghanifard FMM, Zarrinpour V. PYGO2 increases proliferation and migration capacities through critical signaling pathways in esophageal squamous cell carcinoma. J Biochem Mol Toxicol. 2024;38(1):e23625.

    Article  Google Scholar 

  11. Rubie C, et al. Housekeeping gene variability in normal and cancerous colorectal, pancreatic, esophageal, gastric and hepatic tissues. Mol Cell Probes. 2005;19(2):101–9.

    Article  CAS  PubMed  Google Scholar 

  12. Ma B, Hottiger MO. Crosstalk between Wnt/beta-Catenin and NF-kappaB signaling pathway during inflammation. Front Immunol. 2016;7:378.

    Article  PubMed  PubMed Central  Google Scholar 

  13. Gajewski TF, et al. Molecular profiling to identify relevant immune resistance mechanisms in the tumor microenvironment. Curr Opin Immunol. 2011;23(2):286–92.

    Article  CAS  PubMed  Google Scholar 

  14. Wen X, et al. New advances in canonical Wnt/beta-Catenin signaling in Cancer. Cancer Manag Res. 2020;12:6987–98.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Xie YY, et al. Pygo2 regulates adiposity and glucose homeostasis via beta-Catenin-Axin2-GSK3beta signaling pathway. Diabetes. 2018;67(12):2569–84.

    Article  PubMed  Google Scholar 

  16. Andrews PGP, et al. Augmentation of Myc-Dependent mitotic gene expression by the Pygopus2 chromatin effector. Cell Rep. 2018;23(5):1516–29.

    Article  CAS  PubMed  Google Scholar 

  17. Gu B, Watanabe K, Dai X. Pygo2 regulates histone gene expression and H3 K56 acetylation in human mammary epithelial cells. Cell Cycle. 2012;11(1):79–87.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Zhang ZM, et al. Pygo2 activates MDR1 expression and mediates chemoresistance in breast cancer via the Wnt/beta-catenin pathway. Oncogene. 2016;35(36):4787–97.

    Article  CAS  PubMed  Google Scholar 

  19. Liu R, et al. Pygopus 2 promotes kidney cancer OS-RC-2 cells proliferation and invasion in vitro and in vivo. Asian J Urol. 2015;2(3):151–7.

    Article  PubMed  PubMed Central  Google Scholar 

  20. Zhou SY, et al. Overexpression of Pygopus-2 is required for canonical Wnt activation in human lung cancer. Oncol Lett. 2014;7(1):233–8.

    Article  CAS  PubMed  Google Scholar 

  21. Zhou C, et al. Pygo2 functions as a prognostic factor for glioma due to its up-regulation of H3K4me3 and promotion of MLL1/MLL2 complex recruitment. Sci Rep. 2016;6:22066.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Moghbeli M, et al. Association of PYGO2 and EGFR in esophageal squamous cell carcinoma. Med Oncol. 2013;30(2):516.

    Article  PubMed  Google Scholar 

  23. Lu X, et al. An in vivo screen identifies PYGO2 as a driver for metastatic prostate Cancer. Cancer Res. 2018;78(14):3823–33.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Dranoff G. Cytokines in cancer pathogenesis and cancer therapy. Nat Rev Cancer. 2004;4(1):11–22.

    Article  CAS  PubMed  Google Scholar 

  25. Saraiva M, O’Garra A. The regulation of IL-10 production by immune cells. Nat Rev Immunol. 2010;10(3):170–81.

    Article  CAS  PubMed  Google Scholar 

  26. Xin Z, Wenyu F, Shenhua X. Clinicopathologic significance of cytokine levels in esophageal squamous cell carcinoma. Hepatogastroenterology. 2010;57(104):1416–22.

    PubMed  Google Scholar 

  27. Yang WF et al. [Expression of CD80, CD86, TGF-beta1 and IL-10 mRNA in the esophageal carcinoma]. Zhonghua Zhong Liu Za Zhi. 2006;28(10):762-5.

  28. Itakura E, et al. IL-10 expression by primary tumor cells correlates with melanoma progression from radial to vertical growth phase and development of metastatic competence. Mod Pathol. 2011;24(6):801–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Matsumoto K, et al. Interleukin-10 -1082 gene polymorphism and susceptibility to cervical cancer among Japanese women. Jpn J Clin Oncol. 2010;40(11):1113–6.

    Article  PubMed  Google Scholar 

  30. Shi J, et al. IL10 inhibits starvation-induced autophagy in hypertrophic Scar fibroblasts via cross talk between the IL10-IL10R-STAT3 and IL10-AKT-mTOR pathways. Cell Death Dis. 2016;7(3):e2133.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Rutz S, et al. Notch regulates IL-10 production by T helper 1 cells. Proc Natl Acad Sci U S A. 2008;105(9):3497–502.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Gatica-Andrades M, et al. WNT ligands contribute to the immune response during septic shock and amplify endotoxemia-driven inflammation in mice. Blood Adv. 2017;1(16):1274–86.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Hsu TI, et al. Positive feedback regulation between IL10 and EGFR promotes lung cancer formation. Oncotarget. 2016;7(15):20840–54.

    Article  PubMed  PubMed Central  Google Scholar 

  34. Yaguchi T, et al. Immune suppression and resistance mediated by constitutive activation of Wnt/beta-catenin signaling in human melanoma cells. J Immunol. 2012;189(5):2110–7.

    Article  CAS  PubMed  Google Scholar 

  35. Hong Y, et al. Deletion of LRP5 and LRP6 in dendritic cells enhances antitumor immunity. Oncoimmunology. 2016;5(4):e1115941.

    Article  PubMed  Google Scholar 

  36. Zeni E, et al. Macrophage expression of interleukin-10 is a prognostic factor in nonsmall cell lung cancer. Eur Respir J. 2007;30(4):627–32.

    Article  CAS  PubMed  Google Scholar 

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Authors and Affiliations

  1. Department of Clinical Biochemistry, Faculty of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran

    Fereshteh Ahmadian Shalchi

  2. Department of Clinical Biochemistry, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

    Fereshteh Ahmadian Shalchi & Mohammad Arian Ahmadian Shalchi

  3. Division of Human Genetics, Avicenna Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran

    Nayyerehalsadat Hosseini

  4. Department of Biology, Da.C., Islamic Azad University, Cheshmeh-Ali Boulevard, Sa’dei square, P.O.Box: 3671639998, Damghan, Iran

    Fatemeh Nourmohammadi, Mohammad Arian Ahmadian Shalchi & Mohammad Mahdi Forghanifard

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Contributions

F.A.Sh., N.H. and F.N. performed the experiments and drafted the manuscript. M.A.A.Sh. was involved in data analysis and drafting. MMF designed the study, analyzed data, and edited the manuscript.

Corresponding author

Correspondence to Mohammad Mahdi Forghanifard.

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The study was approved by ethics committee of Mashhad University of Medical Sciences (No. 88098) according to the Ethical Helsinki Declaration agreement and informed consent was obtained from all individual participants included in the study.

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Not applicable.

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The authors declare no competing interests.

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Ahmadian Shalchi, F., Hosseini, N., Nourmohammadi, F. et al. PYGO2 regulates IL10 and plays immunosuppressive role through ESCC progression. BMC Mol and Cell Biol 26, 14 (2025). https://doiorg.publicaciones.saludcastillayleon.es/10.1186/s12860-025-00540-0

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  • DOI: https://doiorg.publicaciones.saludcastillayleon.es/10.1186/s12860-025-00540-0

Keywords

BMC Molecular and Cell Biology

ISSN: 2661-8850

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