Research ArticleExperimental Studies
Open Access

Characterization of the Subcellular Distribution of Phospho-β-catenin in Colorectal Cancer

SARP UZUN, AYNUR ISIK, KUBRA KATIPOGLU, GUNES GUNER and AYTEKIN AKYOL
In Vivo July 2023, 37 (4) 1576-1583; DOI: https://doi.org/10.21873/invivo.13242

Abstract

Background/Aim: β-Catenin is a multifunctional protein, which is localized to different subcellular compartments of the normal colon epithelium. The hyperactivation of Wnt pathway results in the nuclear accumulation of β-catenin and induction of colorectal carcinogenesis. Although N-terminally hypo-phosphorylated β-catenin (active β-catenin) is known as the transcriptionally active form, phospho-S33/S37/T41-β-catenin (phospho-β-catenin) can also accumulate in the nucleus. In this study, we aimed to characterize the subcellular distribution of phospho-β-catenin and the other forms of β-catenin in normal colon epithelium and colorectal cancer (CRC). Materials and Methods: Phosphorylated, hypo-phosphorylated, and the total pool of β-catenin were evaluated in colon epithelium and CRC using immunohistochemistry, immunofluorescence staining, and western blotting. Tissue microarrays were used to determine the expression pattern of phospho-β-catenin in CRC samples. Results: Almost 11% (49/452) of CRCs expressed moderate to high levels of phospho-β-catenin in the nucleus. In addition, hypo-phosphorylated and phosphorylated forms of β-catenin localized to different subcellular regions in normal colon epithelium and CRC. Immunoblotting experiments suggested that truncated phospho-β-catenin forms can be found in CRCs. Conclusion: Phospho-β-catenin accumulates in the nucleus and different molecular weight β-catenin proteins are present in colon cancer cells. To elaborate on the functional significance of nuclear phospho-β-catenin, further studies should be performed.

Key Words:

β-Catenin is a multitasking protein that can perform different functions. The majority of the β-catenin is located close to the cell membrane and acts as a molecular bridge between the cytoplasmic domain of E-cadherin and α-catenin. In contrast, a smaller portion of β-catenin shuttles between the cytoplasm and nucleus, where it functions as a transcriptional coactivator of the Wnt pathway (1). Owing to its key role in Wnt signal transduction, the subcellular level and distribution of β-catenin are strictly controlled by a phosphorylation/proteasomal degradation cascade. In the absence of Wnt ligands, cytoplasmic β-catenin forms a complex with APC, Axin, GSK3, and CK1. This complex facilitates the phosphorylation of the N-terminal domain of β-catenin at amino acid residues Ser33, Ser37, Thr41, and Ser45 (2-4). The phosphorylation at Ser33/Ser37 residues creates a motif that is recognized by the E3 ubiquitin ligase β-TrCP and β-catenin is directed to the proteasomal degradation machinery by adding polyubiquitin tags (5). However, the binding of the Wnt ligand to its receptor inhibits N-terminal phosphorylation of β-catenin and its degradation, and β-catenin translocates into the nucleus where it binds to TCF/Lef family of transcription factors and induces the transcription of Wnt target genes (6).

Nuclear accumulation of β-catenin is accepted as an important indicator of active Wnt pathway in colorectal cancer (CRC); however, the phosphorylation status of nuclear β-catenin is the major determinant of Wnt activity as N-terminally hypo-phosphorylated β-catenin, so-called active β-catenin, is intrinsically more efficient at inducing target gene expression (7, 8). N-terminally phosphorylated β-catenin (phospho-β-catenin) has a short half-life compared to the active form and is rapidly degraded by the proteasomal system in the cytoplasm (8). Recent studies on colon cancer cell lines support the notion that both active and phospho-β-catenin localize to different intracellular regions (9-11). However, there is only limited knowledge about how different forms of β-catenin are distributed in tissue samples.

Therefore, we used different antibodies raised against N-terminally phosphorylated and/or unphosphorylated forms of β-catenin and evaluated the expression pattern on tissue microarrays (TMAs) constructed using formalin-fixed and paraffin-embedded (FFPE) CRC patient samples. In addition to FFPE tissue samples, fresh frozen normal colon and CRC patient samples were analyzed using immunofluorescence staining and immunoblotting. With this study, we examined the subcellular distribution of phospho-β-catenin and the other molecular forms of β-catenin in normal colon epithelium and primary human CRC samples.

Materials and Methods

Tissue samples and colorectal cancer tissue microarrays (TMA). A total of 19 CRC tissue microarrays representing 472 CRC patients were used in this study. All FFPE tissue samples were retrieved from the archives of Hacettepe University Department of Pathology. Four of the 19 TMAs (TMA 1-4) were previously constructed using FFPE tissue blocks of CRC patients. These TMAs were composed of duplicate tissue cores and contained a total of 203 CRC samples with various clinicopathological features (12). The other group of TMAs (TMA 5-19) was composed of two 3 mm diameter tissue cores from each CRC patient, one of the cores from the periphery of the tumor and the other one from the center of the tumor. These TMAs represent a total of 269 CRC patients. To perform immunofluorescence staining and western blotting, patient samples from normal colon and colorectal cancer tissues were also collected in addition to FFPE samples. The patient selection process and study design were executed according to the regulations of the Non-Interventional Clinical Research Ethics Committee of Hacettepe University (GO 18/485-18) and the Declaration of Helsinki.

Immunohistochemistry. Four-μm-thick tissue sections were acquired from TMAs. For deparaffinization, sections were incubated at 75°C for 45 min and soaked into xylene, then rehydrated in series of decreasing alcohol concentrations. Endogenous hydrogen peroxide activity was inhibited with 7% H202/80% methanol solution. Heat-induced epitope retrieval was performed using a pressure cooker in citrate buffer (pH: 6.0) or EDTA buffer (pH: 8.0). Sections were incubated overnight at 4°C with three different β-catenin antibodies: Mouse monoclonal anti-β-catenin [1:1,000] (610154, BD Biosciences, San Jose, CA, USA) antibody (BC antibody), which recognizes the intracellular pool of β-catenin by binding to its C-terminal region; mouse monoclonal anti-active-β-catenin clone 8E7 [1:100] (05-665, Merck Millipore, Burlington, MA, USA) antibody (ABC antibody), which was raised against the N-terminal region and can specifically recognize β-catenin that is unphosphorylated at S37 and Thr41 amino acid residues; rabbit polyclonal phospho-β-catenin (Ser33/Ser37/Thr41) [1:50] (9561, Cell Signaling Technology, Beverly, MA, USA) antibody (PBC antibody), which recognizes β-catenin when Ser33, Ser37, and Thr41 amino acid residues are phosphorylated. After incubation with primary antibodies, sections were rinsed with TBS and incubated with biotinylated goat anti-polyvalent secondary antibody and streptavidin peroxidase (Lab Vision, anti-polyvalent HRP; Thermo Fisher Scientific, Waltham, MA, USA). Subsequently, sections were incubated with the chromogen 3,3′-diaminobenzidine (DAB, Thermo Fischer Scientific) and counterstained with hematoxylin. All TMAs were scanned with Olympus VS 120 System (Olympus, Tokyo, Japan) and visualized with OlyVIA software (Olympus).

Immunohistochemical staining pattern and intensity were evaluated as previously described (13). Briefly, each subcellular area (membrane, cytoplasm, and nucleus) was evaluated separately for each sample. Staining intensity was scored as follows: no staining 0, weak staining 1, moderate staining 2, and strong staining 3. Samples that had regions with different staining intensities were scored according to the predominant staining, Each sample was evaluated by two observers with two-fold redundancy. All antibodies were used according to manufacturers’ validation data and the antibody dilutions for each application are summarized in Table I.

View this table:
Table I.

β-Catenin antibody list.

Immunofluorescence staining. Fresh frozen tissues of normal colon and colon adenocarcinomas were cut into 8-μm-thick sections and fixed in 100% methanol for 15 min. Fixed sections were incubated with anti-β-catenin antibody (1:1,000) and phospho-β-catenin (Ser33/37/Thr41) antibody (1:50) overnight at 4°C. For secondary antibodies; Goat Anti-Mouse IgG H&L (Alexa Fluor® 647, 1:200, ab150115, Abcam, Cambridge, MA, USA) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488, 1:1,000, ab150077, Abcam) were applied for 1 h at room temperature. Nuclei were stained with DAPI (4′,6-diamidino-2-phenylindole). Images were captured using Leica DM2500 (Leica Camera, Wetzlar, Germany).

Western blot analysis. Whole-cell lysates of normal colon epithelium and colon adenocarcinoma were prepared in RIPA Buffer containing cOmplete™, Mini, EDTA-free Protease Inhibitor Cocktail (Sigma-Aldrich, St. Louis, MO, USA). Proteins were separated with gel electrophoresis and transferred to PVDF membranes. Membranes were incubated overnight at 4°C with anti-β-catenin antibody [1:2,000] (05-665, Merck Millipore), non-phospho (Active) β-catenin (Ser33/37/Thr41) (D13A1) Rabbit mAb [1: 1.000] (8814, Cell Signaling Technology, Beverly, MA, USA) and phospho-β-catenin (Ser33/37/Thr41) antibody [1:500] (9561, Cell Signaling Technology). After washing steps, membranes were incubated for 1 h at room temperature with appropriate secondary antibodies and developed using ECL solution (32106, Thermo Fisher Scientific). For loading control, anti-β-Actin-Peroxidase [1:30,000] (A3854, Sigma-Aldrich) was used. Images were obtained with FluorChem FC3 system (Protein Simple, San Jose, CA, USA).

Statistical analysis. Descriptive analyses are presented as percentage and frequency tables for categorical variables and as median and minimum-maximum values for numerical variables having non-normal distribution. Survival curves were generated with the Kaplan-Meier method and the curves were compared for time-to-event measures with the log-rank test. We defined survival as the time from CRC diagnosis to death from any cause. p-Values <0.05 were considered statistically significant. Statistical analyses were performed using GraphPad Prism version 9.2.0 for Windows (GraphPad Software, San Diego, CA, USA). All figures were created with Inkscape version 1.1.1 (Inkscape Project).

Results

Immunohistochemical evaluation of phospho-β-catenin in colorectal cancer samples. We first stained all TMAs with the PBC antibody. Out of 472, we were able to assess phospho-β-catenin expression in 452 sections from CRC patients due to technical reasons such as loss of tissue cores during heat-induced epitope retrieval. Phospho-β-catenin was either expressed inside the nucleus or cytoplasm of neoplastic cells and only nine cases had both nuclear and cytoplasmic staining. Of all cases, 25.8% (55/452) showed nuclear staining with phospho-β-catenin and 10.8% (49/452) showed moderate to strong staining. Cytoplasmic staining was only detected in 18.8% (85/452) of the cases and was faint in most cases. In contrast, more than 50% of all cases had neither nuclear nor cytoplasmic staining (Figure 1A-C). Normal colon and colon adenoma samples were also probed with the PBC antibody; only poor staining was observed in few cases (data not shown). The clinicopathologic features of the CRC cases are summarized in Table II.

Figure 1.
Figure 1.
Figure 1.

Immunohistochemistry, immunofluorescence, and immunoblotting of β-catenin using different antibodies in colorectal cancer specimens. Colorectal cancer samples showed three different phospho-β-catenin staining patterns: nuclear staining (A), cytoplasmic staining (B), and negative staining (C). The black arrow shows nuclear accumulation, and the red arrow shows the cytoplasmic accumulation of phospho-β-catenin. Scale bars, 50 μm. The same CRC tissue sample was stained with different β-catenin antibodies: Phospho-β-catenin stained only the nucleus (D), the BC antibody-stained nucleus, cytoplasm, and membrane (E). The ABC antibody mainly stained the membrane and to a lesser extent the cytoplasm (F). Black arrows show nuclear accumulation of phospho-β-catenin and β-catenin in D and E, respectively. Scale bars, 50 μm. Immunofluorescence staining of a normal colon sample with phospho-β-catenin and anti-β-catenin antibodies. Arrows indicate the punctate cytoplasmic staining of phospho-β-catenin. Arrowhead shows phospho-β-catenin-expressing endothelial cells (G). Immunofluorescence staining of the colorectal cancer sample with PBC and BC antibodies. Different antibodies stain different subcellular compartments. Arrowhead shows the phospho-β-catenin expressed endothelial cells (H). Immunoblotting of protein lysates from colorectal cancer samples with phospho-β-catenin, anti-β-catenin, and anti-ABC antibodies. The black arrow indicates the expected location of β-catenin band. Red arrows indicate bands/expected bands of cleaved β-catenin without N-terminal. The blue arrow indicates Ig heavy chain (I).

View this table:
Table II.

Clinicopathological features of 452 colorectal cancer samples in tissue microarrays.

Evaluation of different forms of β-catenin with immunohistochemistry, immunofluorescence staining and immunoblotting. In order to evaluate the phospho-β-catenin expression and its subcellular association with the other forms of β-catenin, we collected normal and cancer tissues from CRC patients and first performed immunohistochemical staining with PBC, BC, and ABC antibodies. In cases having no expression of phospho-β-catenin; the BC and ABC antibodies stained the membrane, cytoplasm, and nucleus of the tumor cells. In cases expressing nuclear phospho-β-catenin, the BC antibody again stained all compartments of the tumor cells (Figure 1D and E). However, when these same cases were stained with the ABC antibody, nuclei of neoplastic cells could not be stained although active-β-catenin expression was detected on the membrane (Figure 1F). Only the membrane of normal colon epithelium was stained with BC and ABC antibodies, as expected (data not shown).

To investigate the distribution of β-catenin and phospho-β-catenin at the cellular level, we performed immunofluorescence staining using the BC and PBC antibodies. We showed that the BC antibody diffusely stained membranes of normal colon epithelial cells (Figure 1G). However, phospho-β-catenin accumulated inside the apical cytoplasm of the colon epithelium and gave a punctate staining pattern (arrow). In contrast to normal tissue, phospho-β-catenin was strictly localized to nuclei of neoplastic cells of CRC and similarly to normal tissue, a punctate staining pattern was observed (Figure 1H). Interestingly, phospho-β-catenin expression was not only limited to colon epithelium but also detected in endothelial cells of both normal and neoplastic tissues (arrowhead).

In normal colon epithelium and some of the colon cancer cells, β-catenin staining was limited to the membrane and did not colocalize with phospho-β-catenin. Since BC and PBC antibodies were raised against two different terminals of β-catenin, we thought that there might be C-terminal truncated β-catenin forms. To examine this hypothesis, protein isolates from CRC cases were assessed using immunoblotting with three different β-catenin antibodies (Figure 1I). The PBC antibody revealed three extra bands between 60 kDa and 120 kDa. The expected β-catenin band gave no to very weak signal (black arrow) that became visible only in overexposed images (data not shown). The BC antibody, however, revealed four different bands between 75 kDa and 110 kDa, but surprisingly, two low molecular weight bands disappeared after immunoblotting of the same membrane with the ABC antibody (red arrows). Immunoblotting with both antibodies revealed the expected β-catenin band at 95 kDa and an extra band at approximately 110 kDa.

Effect of phospho-β-catenin expression on overall survival of colorectal cancer patients. We first categorized nuclear and cytoplasmic phospho-β-catenin staining as follows: no staining (0) represented the phospho-β-catenin negative group, and weak to strong staining (≥1) represented the phospho-β-catenin positive group. We could not display any effect of nuclear (p=0.328) or cytoplasmic (p=0.192) phospho-β-catenin expression on the overall survival of patients with CRC (Figure 2A and B). Then, we changed our categorization system and patients were divided into two groups according to nuclear phospho-β-catenin staining as follows: group 1, no to weak staining (0-1) and group 2 moderate to strong staining (2-3). Although moderate to strong phospho-β-catenin expression seemed to create predilection towards poor survival, we didn’t find a statistically significant association with overall survival (p=0.093) (Figure 2C). In addition, we also evaluated whether other variables had an impact on survival of patients with CRC. In our cohort, we could only find a significant association between tumor stage and survival (p<0.0001), but we could not show the effect of sex (p=0.396) or tumor location (p=0.633) on overall survival.

Figure 2.
Figure 2.

Effect of phospho-β-catenin expression on overall survival of patients with colorectal cancer. Kaplan-Meier analysis of the effect of nuclear phospho-β-catenin on overall survival of patients with colorectal cancer. Orange and blue curves represent no staining (0) and weak to strong staining (≥1), respectively (A). Kaplan-Meier analysis of the effect of cytoplasmic phospho-β-catenin on overall survival of patients with colorectal cancer. Orange and blue curves represent no staining (0) and weak to strong staining (≥1), respectively (B). Kaplan-Meier analysis of the effect of nuclear phospho-β-catenin on overall survival of patients with colorectal cancer. Orange and blue curves represent no to weak staining (0-1) and moderate to strong staining (2-3), respectively (C).

Discussion

β-Catenin has multiple functions in almost every compartment of the cell. As the central component of the canonical WNT pathway, its level and intracellular distribution are strictly regulated with a phosphorylation-ubiquitination-proteasomal degradation cascade. Phospho-β-catenin is rapidly degraded in the cytoplasm by the 26S proteasome complex and therefore has a short half-life. However, some studies have suggested that the localization of phospho-β-catenin is not limited to the cytoplasm, but it may also accumulate in the nucleus and membrane (9, 14).

In this study, we observed that in approximately 25% of CRC cases phospho-β-catenin was expressed in the nucleus and 11% of the cases showed moderate to strong staining, a rate slightly higher than that reported previously (13). In contrast to this previous study, cytoplasmic expression of phospho-β-catenin was also observed in some cases and the staining was mainly of weak to moderate intensity. In a study on invasive breast carcinomas, nuclear phospho-β-catenin was associated with an aggressive tumor phenotype, whereas cytoplasmic localization was linked to a more favorable phenotype (15). In our study, we did not observe any association between the localization of phospho-β-catenin and CRC tumor phenotype. Since nuclear staining almost never coincided with cytoplasmic staining in our cases, we thought that cytoplasmic accumulation of phospho-β-catenin may indicate β-catenin dysregulation and may underlie a different pathogenetic mechanism.

We and another group have described an immunohistochemical method that can detect CTNNB1 exon 3 mutations by using two different antibodies raised against the C-terminal and N-terminal of β-catenin (12, 16). Since exon 3 mutations cause alterations in a hotspot region at the N-terminal, antibodies raised against this specific region cannot bind the mutated form of β-catenin and give no to weak signal although the C-terminal antibody shows strong nuclear staining. As shown in Figure 1E and F, we observed a similar staining pattern in some cases; nuclear staining with the C-terminal antibody (BC antibody) but no nuclear staining with the N-terminal antibody (ABC antibody). Interestingly, the same CRC samples gave strong nuclear staining with phospho-β-catenin antibody as shown in Figure 1D. Therefore, we suggested that the ABC antibody cannot detect phospho-β-catenin and leads to a staining pattern similar to that observed in cases having CTNNB1 exon 3 mutation in our previous study.

There are a few studies that investigated the potential intracellular functions of phospho-β-catenin. Using a rat fibroblast cell line, Huang et al. first showed that phospho-β-catenin was clustered in centrosomes and regulated both microtubule organization and centrosome maturation (17). In a following study, centrosomal localization of phospho-β-catenin was verified and it was shown that phospho-β-catenin was also responsible for centrosome cohesion (18). Although there is strong evidence that phospho-β-catenin is clustered in centrosomes to regulate the organization of microtubules, there is no information to explain by which biochemical mechanisms it carries out this function (19). Localization of phospho-β-catenin specifically to the centrosomes and its involvement in microtubule organization may suggest a potential role of nuclear phospho-β-catenin in cell cycle regulation.

In our study, we also evaluated the intracellular distribution of phospho-β-catenin and β-catenin using immunofluorescence staining. Interestingly, β-catenin was not detected in the nucleus of some neoplastic cells and did not colocalize with phospho-β-catenin. One possible explanation is that β-catenin is present in different isoforms, some of which are truncated and cannot be recognized by the BC antibody. In support, immunoblotting with phospho-β-catenin revealed approximately 60 kDa and 80 kDa low molecular weight bands. It is well known that β-catenin frequently undergoes posttranslational modifications and the most common of these is the proteolytic cleavage of the protein. Goretsky et al. (20) have recently suggested that β-catenin can be cleaved from its C-terminal and N-terminal regions by the chymotrypsin-like activity of the proteasomal system, which leads to the production of low molecular weight β-catenin. Therefore, it is possible that we observed low molecular weight bands without a C-terminal and detected almost no signal at 92 kDa due to the massive production of truncated β-catenin.

After immunoblotting with the BC antibody, we also observed two extra, low molecular weight bands which disappeared after immunoblotting with the ABC antibody. Several studies suggested that β-catenin can be cleaved by the calpain family of proteases from its N-terminal region and becomes transcriptionally active due to the lack of phosphorylation motif. A 2002 study showed that a 75 kDa β-catenin band appears after immunoblotting of protein lysates of colorectal cell lines and the same β-catenin band was also detected in a following study on prostate and breast cancer cell lines (21, 22). In these studies, a β-catenin band around 90 kDa was also detected which indicated that the low molecular weight bands that we observed in our study might be the products of N-terminal cleavage.

A previous study showed that phospho-β-catenin expression was associated with better survival in CRC patients (13). However, in the present study, we could not find any effect of the nuclear or cytoplasmic phospho-β-catenin expression on the overall survival of CRC patients. Therefore, the prognostic value of phospho-β-catenin expression in CRC should be reconsidered in further studies.

In conclusion, we evaluated the subcellular distribution of different isoforms of β-catenin using antibody-based staining approaches and showed that phospho-β-catenin can accumulate in the nuclei of neoplastic cells of CRC. Furthermore, immunoblotting analysis indicated the presence of multiple β-catenin bands in CRC, which may be different β-catenin isoforms that derive from proteolytic processing. Further investigation will elaborate on the biological functions and the role of these different molecular weight β-catenin isoforms and provide a better understanding of CRC pathogenesis.

Footnotes

  • ↵* Present address: Department of Pathology, Ankara City Hospital, Ankara, Turkey.

  • ↵# Present address: The Institute of Medical Genetics and Pathology, University Hospital Basel, Basel, Switzerland.

  • Authors’ Contributions

    S.U., A.I., and K.K. carried out the experiments. G.G. and A.A. supervised S.U. S.U. and A.A. wrote the manuscript. All Authors discussed the results and contributed to the final manuscript.

  • Conflicts of Interest

    The Authors have no conflicts of interest to declare in relation to this study.

  • Received April 19, 2023.
  • Revision received May 10, 2023.
  • Accepted May 11, 2023.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY-NC-ND) 4.0 international license (https://creativecommons.org/licenses/by-nc-nd/4.0).

References

  1. ↵
    1. Valenta T,
    2. Hausmann G,
    3. Basler K
    : The many faces and functions of β-catenin. EMBO J 31(12): 2714-2736, 2022. DOI: 10.1038/emboj.2012.150
    OpenUrlCrossRef
  2. ↵
    1. Yost C,
    2. Torres M,
    3. Miller J,
    4. Huang E,
    5. Kimelman D,
    6. Moon R
    : The axis-inducing activity, stability, and subcellular distribution of beta-catenin is regulated in Xenopus embryos by glycogen synthase kinase 3. Genes Dev 10(12): 1443-1454, 2021. DOI: 10.1101/gad.10.12.1443
    OpenUrlCrossRef
    1. Liu C,
    2. Li Y,
    3. Semenov M,
    4. Han C,
    5. Baeg G,
    6. Tan Y,
    7. Zhang Z,
    8. Lin X,
    9. He X
    : Control of β-catenin phosphorylation/degradation by a dual-kinase mechanism. Cell 108(6): 837-847, 2021. DOI: 10.1016/s0092-8674(02)00685-2
    OpenUrlCrossRef
  4. ↵
    1. Amit S,
    2. Hatzubai A,
    3. Birman Y,
    4. Andersen J,
    5. Ben-Shushan E,
    6. Mann M,
    7. Ben-Neriah Y,
    8. Alkalay I
    : Axin-mediated CKI phosphorylation of β-catenin at Ser 45: a molecular switch for the Wnt pathway. Genes Dev 16(9): 1066-1076, 2021. DOI: 10.1101/gad.230302
    OpenUrlCrossRef
  5. ↵
    1. Aberle H,
    2. Bauer A,
    3. Stappert J,
    4. Kispert A,
    5. Kemler R
    : beta-catenin is a target for the ubiquitin-proteasome pathway. EMBO J 16: 3797-3804, 1997. DOI: 10.1093/emboj/16.13.3797
    OpenUrlAbstract
  6. ↵
    1. Archbold H,
    2. Yang Y,
    3. Chen L,
    4. Cadigan K
    : How do they do Wnt they do?: regulation of transcription by the Wnt/β-catenin pathway. Acta Physiol 204(1): 74-109, 2021. DOI: 10.1111/j.1748-1716.2011.02293.x
    OpenUrlCrossRef
  7. ↵
    1. Van Noort M,
    2. Meeldijk J,
    3. Van der Zee R,
    4. Destree O,
    5. Clevers H
    : Wnt signaling controls the phosphorylation status of β -catenin. J Biol Chem 277(20): 17901-17905, 2022. DOI: 10.1074/jbc.M111635200
    OpenUrlCrossRef
  8. ↵
    1. Staal F,
    2. Van noort M,
    3. Strous G,
    4. Clevers H
    : Wnt signals are transmitted through N-terminally dephosphorylated beta-catenin. EMBO Rep 3(1): 63-68, 2021. DOI: 10.1093/embo-reports/kvf002
    OpenUrlCrossRef
  9. ↵
    1. Sadot E,
    2. Conacci-Sorrell M,
    3. Zhurinsky J,
    4. Shnizer D,
    5. Lando Z,
    6. Zharhary D,
    7. Kam Z,
    8. Ben-Ze’ev A,
    9. Geiger B
    : Regulation of S33/S37 phosphorylated beta-catenin in normal and transformed cells. J Cell Sci 115: 2771-2780, 2002.
    OpenUrlAbstract/FREE Full Text
    1. Maher MT,
    2. Mo R,
    3. Flozak AS,
    4. Peled ON,
    5. Gottardi CJ
    : Beta-catenin phosphorylated at serine 45 is spatially uncoupled from beta-catenin phosphorylated in the GSK3 domain: implications for signaling. PLoS One 5: e10184, 2010. DOI: 10.1371/journal.pone.0010184
    OpenUrlCrossRefPubMed
  11. ↵
    1. Faux M,
    2. Coates J,
    3. Kershaw N,
    4. Layton M,
    5. Burgess A
    : Independent interactions of phosphorylated β-catenin with E-cadherin at cell-cell contacts and APC at cell protrusions. PLoS One 5(11): e14127, 2020. DOI: 10.1371/journal.pone.0014127
    OpenUrlCrossRef
  12. ↵
    1. Akyol A,
    2. Güner G,
    3. Özşeker HS,
    4. Işık A,
    5. Atcı Ö,
    6. Uzun S,
    7. Atayar E,
    8. Ozaltin F,
    9. Gedikoğlu G,
    10. Sökmensüer C,
    11. Fearon ER
    : An immunohistochemical approach to detect oncogenic CTNNB1 mutations in primary neoplastic tissues. Lab Invest 99: 128-137, 2019. DOI: 10.1038/s41374-018-0121-9
    OpenUrlCrossRefPubMed
  13. ↵
    1. Chung GG,
    2. Provost E,
    3. Kielhorn EP,
    4. Charette LA,
    5. Smith BL,
    6. Rimm DL
    : Tissue microarray analysis of beta-catenin in colorectal cancer shows nuclear phospho-beta-catenin is associated with a better prognosis. Clin Cancer Res 7: 4013-4020, 2001.
    OpenUrlAbstract/FREE Full Text
  14. ↵
    1. Maher MT,
    2. Flozak AS,
    3. Stocker AM,
    4. Chenn A,
    5. Gottardi CJ
    : Activity of the β-catenin phosphodestruction complex at cell–cell contacts is enhanced by cadherin-based adhesion. J Cell Biol 186: 219-228, 2009. DOI: 10.1083/jcb.200811108
    OpenUrlAbstract/FREE Full Text
  15. ↵
    1. Nakopoulou L,
    2. Mylona E,
    3. Papadaki I,
    4. Kavantzas N,
    5. Giannopoulou I,
    6. Markaki S,
    7. Keramopoulos A
    : Study of phospho-beta-catenin subcellular distribution in invasive breast carcinomas in relation to their phenotype and the clinical outcome. Mod Pathol 19: 556-563, 2006. DOI: 10.1038/modpathol.3800562
    OpenUrlCrossRefPubMed
  16. ↵
    1. Dubbink H,
    2. Hollink I,
    3. Avenca Valente C,
    4. Wang W,
    5. Liu P,
    6. Doukas M,
    7. Van Noesel M,
    8. Dinjens W,
    9. Wagner A,
    10. Smits R
    : A novel tissue-based β-catenin gene and immunohistochemical analysis to exclude familial adenomatous polyposis among children with hepatoblastoma tumors. Pediatr Blood Cancer 65(6): e26991, 2018. DOI: 10.1002/pbc.26991
    OpenUrlCrossRef
  17. ↵
    1. Huang P,
    2. Senga T,
    3. Hamaguchi M
    : A novel role of phospho-beta-catenin in microtubule regrowth at centrosome. Oncogene 26(30): 4357-4371, 2022. DOI: 10.1038/sj.onc.1210217
    OpenUrlCrossRef
  18. ↵
    1. Hadjihannas M,
    2. Brückner M,
    3. Behrens J
    : Conductin/axin2 and Wnt signalling regulates centrosome cohesion. EMBO Rep 11(4): 317-324, 2021. DOI: 10.1038/embor.2010.23
    OpenUrlCrossRef
  19. ↵
    1. Mbom BC,
    2. Nelson WJ,
    3. Barth A
    : β-catenin at the centrosome: Discrete pools of β-catenin communicate during mitosis and may co-ordinate centrosome functions and cell cycle progression. BioEssays 35: 804-809, 2013. DOI: 10.1002/bies.201300045
    OpenUrlCrossRefPubMed
  20. ↵
    1. Goretsky T,
    2. Bradford E,
    3. Ye Q,
    4. Lamping O,
    5. Vanagunas T,
    6. Moyer M,
    7. Keller P,
    8. Sinh P,
    9. Llovet J,
    10. Gao T,
    11. She Q,
    12. Li L,
    13. Barrett T
    : Beta-catenin cleavage enhances transcriptional activation. Sci Rep 8(1): 671, 2022. DOI: 10.1038/s41598-017-18421-8
    OpenUrlCrossRef
  21. ↵
    1. Li G,
    2. Iyengar R
    : Calpain as an effector of the Gq signaling pathway for inhibition of Wnt/beta-catenin-regulated cell proliferation. Proc Natl Acad Sci USA 99(20): 13254-13259, 2022. DOI: 10.1073/pnas.202355799
    OpenUrlCrossRef
  22. ↵
    1. Rios-Doria J,
    2. Kuefer R,
    3. Ethier S,
    4. Day M
    : Cleavage of beta-catenin by calpain in prostate and mammary tumor cells. Cancer Res 64(20): 7237-7240, 2022. DOI: 10.1158/0008-5472.CAN-04-1048
    OpenUrlCrossRef
Back to top

In this issue

Print
Download PDF
Article Alerts
Email Article
Citation Tools
Reprints and Permissions
Characterization of the Subcellular Distribution of Phospho-β-catenin in Colorectal Cancer
SARP UZUN, AYNUR ISIK, KUBRA KATIPOGLU, GUNES GUNER, AYTEKIN AKYOL
In Vivo Jul 2023, 37 (4) 1576-1583; DOI: 10.21873/invivo.13242
Twitter logo Facebook logo Mendeley logo

Jump to section

Keywords