ILK expression in human basal cell carcinoma correlates with epithelial–mesenchymal transition markers and tumour invasion

Papanikolaou S, Bravou V, Gyftopoulos K, Nakas D, Repanti M & Papadaki H
(2010) Histopathology 56 , 799–809
ILK expression in human basal cell carcinoma correlates with epithelial–mesenchymal transition markers and tumour invasion Aims: Epithelial–mesenchymal transition (EMT) has been known to play a significant role in tumour progression. Integrin‐linked kinase (ILK) has been recently added to the growing list of EMT regulators that control some aspect of carcinogenesis. The aim was to study ILK expression and its relevance to EMT markers in human basal cell carcinoma (BCC). Methods and results: Paraffin‐embedded tissue sections from 100 human BCC cases were processed by immunohistochemistry for the expression of ILK, E‐cadherin, Snail, β‐catenin and alpha‐smooth muscle actin (α‐SMA). ILK overexpression was observed in 100% of cases and strongly correlated with tumour invasion and infiltrative BCC. Loss of membranous E‐cadherin was found in 71% of cases while nuclear immunoreactivity for E‐cadherin was also observed in 90% of the tumours. Snail, nuclear β‐catenin and α‐SMA expression was detected in 100%, 99% and 97% of tumours, respectively. Aberrant expression of E‐cadherin, nuclear β‐catenin and α‐SMA correlated with BCC tumour invasion. Interestingly, there was a significant correlation between ILK expression and all the EMT markers examined. Conclusions: ILK overexpression in BCC is implicated in tumour progression probably through the induction of an EMT‐related molecular profile. Nuclear localization of E‐cadherin in BCC is also associated with aggressive tumour features.     

Involvement of α‐ and β‐catenins and E‐cadherin in the development of mammary phyllodes tumours

Tsang J Y S, Mendoza P, Lam C C F, Yu A M C, Putti T C, Karim R Z, Scolyer R A, Lee C S, Tan P H & Tse G M
(2012) Histopathology  61, 667–674 Involvement of α‐ and β‐catenins and E‐cadherin in the development of mammary phyllodes tumours Aims: Phyllodes tumours (PT) are rare but clinically important fibroepithelial tumours of the breast. β‐Catenin, a key component in Wnt signalling, has been shown to be important in the development of PT. It also functions as a component of the cadherin complex, which may therefore be implicated in PT pathogenesis. By assessing stromal α‐catenin, β‐catenin and E‐cadherin expression in 158 PT cases using immunohistochemistry and examining associations with clinicopathological features, we aimed to determine the role of these proteins in PT pathogenesis. Methods and results: Cytoplasmic β‐catenin correlated with α‐catenin expression. A significantly higher expression of both markers was observed in borderline than in benign PT (P = 0.003 and <0.001, respectively), but a lower level was found in malignant PT. Cytoplasmic E‐cadherin expression was significantly higher in borderline and malignant than in benign PT (P = 0.001 and 0.012, respectively), but was not correlated with other markers. Both E‐cadherin and α‐catenin showed stronger correlations with histological parameters than β‐catenin. α‐Catenin showed a significant correlation with recurrence (P = 0.005 and 0.016, respectively). Conclusions: α‐ and β‐catenins may be important in the early stages of PT development, while E‐cadherin may be required for malignant development. The correlation of α‐catenin expression with tumour recurrence may be relevant in predicting PT behaviour.     

Nuclear translocation of β-catenin synchronized with loss of E-cadherin in oral epithelial dysplasia with a characteristic two-phase appearance

Alvarado C G, Maruyama S, Cheng J, Ida‐Yonemochi H, Kobayashi T, Yamazaki M, Takagi R & Saku T
(2011) Histopathology 59 , 283–291 Nuclear translocation of β‐catenin synchronized with loss of E‐cadherin in oral epithelial dysplasia with a characteristic two‐phase appearance Aims: One of the important histopathological characteristics of oral epithelial dysplasia is a two‐phase appearance of rete processes, comprising an upper layer of keratinized cells and a lower layer of basaloid cells, and thereby creating a sharp contrast between these two separate cell populations. The aim of this study was to determine the cellular adhesion status of the basaloid cells. Methods and results: Immunohistochemistry for β‐catenin, E‐cadherin and their related molecules was carried out in surgical specimens of two‐phase epithelial dysplasia of the oral mucosa. The lower‐half basaloid cells and the upper keratinized cells were microdissected separately, and extracted DNA samples were subjected to methylation‐specific polymerase chain reaction amplification for E‐cadherin. β‐Catenin was immunolocalized within the nuclei and cytoplasm of Ki67‐positive lower‐half basaloid cells, as well as on the cell membrane of upper parakeratotic cells. The basaloid cells of the lower‐half were also positive for matrix metalloproteinase‐7 and cyclin D1, β‐catenin target gene products, α‐dystroglycan, tenascin‐C, and perlecan, but not for E‐cadherin. The promoter region of the E‐cadherin gene was hypermethylated. Conclusions: The solid proliferation of lower‐half E‐cadherin‐free basaloid cells is enhanced by Wnt signalling cascades, as well as by the intraepithelial extracellular matrix or its bound growth factors.     

Stromal CD10 expression and relationship to the E‐cadherin/β‐catenin complex in breast carcinoma

Kim H‐S, Kim G Y, Kim Y W, Park Y‐K, Song J‐Y & Lim S‐J
(2010) Histopathology 56, 708–719
Stromal CD10 expression and relationship to the E‐cadherin/β‐catenin complex in breast carcinoma Aims: Previous investigations have indicated that stromal CD10 expression, and altered levels of both E‐cadherin and β‐catenin, are associated with the biological aggressiveness of human carcinoma. The aim was to evaluate stromal CD10 expression and the association of stromal CD10 with E‐cadherin and β‐catenin in breast carcinoma. Methods and results: The expression of CD10, E‐cadherin and β‐catenin was immunohistochemically analysed in tissue microarrays containing 104 cases of invasive ductal carcinoma (IDC) and 10 cases of ductal carcinoma in situ (DCIS). Stromal CD10 was detected in 49.5% (50/101) of the IDC. No immunoreactivity was identified in the stromal cells of normal breast, DCIS or intraductal components of IDC. Accumulation of the cytoplasmic β‐catenin was found in 87.0% (87/100) of the IDC. Stromal CD10 expression in IDC was significantly correlated with tumour size (P = 0.027), stage (P < 0.001) and histological grade (P = 0.006), the presence of nodal (P = 0.048) and distant (P = 0.015) metastases, oestrogen receptor‐negative status (P = 0.016), cytoplasmic β‐catenin accumulation (P = 0.031) and lower overall survival rate (P = 0.041). Conclusions: Stromal CD10 expression in IDC may constitute an important prognostic marker. Stromal CD10 expression with associated aggressive features might be related to aberrant β‐catenin expression.     

Anti-Müllerian hormone inhibits growth of AMH type II receptor-positive human ovarian granulosa cell tumor cells by activating apoptosis

Ovarian granulosa cell tumors (GCTs) are sex cord stromal tumors that constitute 3-5% of all ovarian cancers. GCTs usually present with an indolent course but there is a high risk of recurrence, which associates with increased mortality, and targeted treatments would be desirable. Anti-Müllerian hormone (AMH), a key factor regulating sexual differentiation of the reproductive organs, has been implicated as a growth inhibitor in ovarian cancer. GCTs and normal granulosa cells produce AMH, but its expression in large GCTs is usually downregulated. Further, as the lack of specific AMH-signaling pathway components leads to GCT development in mice, we hypothesized that AMH inhibits growth of GCTs. Utilizing a large panel of human GCT tissue samples, we found that AMH type I receptors (ALK2, ALK3 and ALK6) and type II receptor (AMHRII), as well as their downstream effectors Smad1/5, are expressed and active in GCTs. AMHRII expression was detected in the vast majority (96%) of GCTs and correlated with AMH mRNA and protein expression. AMH mRNA level was low in large GCTs, confirming previous findings on low-AMH protein expression in large human as well as mouse GCTs. To study the functional role of AMH in this peculiar ovarian cancer, we utilized a human GCT cell line (KGN) and 10 primary GCT cell cultures. We found that the AMH-Smad1/5-signaling pathway was active in these cells, and that exogenous AMH further activated Smad1/5 in KGN cells. Furthermore, AMH treatment reduced the number of KGN cells and primary GCT cells, with increasing amounts of AMH leading to augmented activation of caspase-3 and subsequent apoptosis. All in all, these data support the premise that AMH is a growth inhibitor of GCTs.     

Nuclear translocation of β‐catenin and decreased expression of epithelial cadherin in human papillomavirus‐positive tonsillar cancer: an early event in human papillomavirus‐related tumour progression?

Stenner M, Yosef B, Huebbers C U, Preuss S F, Dienes H‐P, Speel E‐J M, Odenthal M & Klussmann J P (2011) Histopathology 58 , 1117–1126 Nuclear translocation of β‐catenin and decreased expression of epithelial cadherin in human papillomavirus‐positive tonsillar cancer: an early event in human papillomavirus‐related tumour progression? Aims: High‐risk human papillomaviruses (HPVs) constitute an important risk factor for tonsillar cancer. This study describes changes in cell adhesion molecules during metastasis of HPV‐related and HPV‐unrelated tonsillar carcinomas. Methods and results: We examined 48 primary tonsillar carcinoma samples (25 HPV‐16 DNA‐positive, 23 HPV‐16 DNA‐negative) and their respective lymph node metastases for their HPV status and for the expression of p16, epithelial cadherin (E‐cadherin), β‐catenin, and vimentin. A positive HPV‐specific polymerase chain reaction finding correlated significantly with p16 overexpression in both primary tumours and their metastases (P < 0.0001 for both). In HPV‐unrelated carcinomas, the expression of E‐cadherin was significantly lower in metastases than in primary tumours (P < 0.001). In contrast, the expression of nuclear β‐catenin was significantly higher in metastases than in primary tumours (P = 0.016). In HPV‐related carcinomas, nuclear localization of β‐catenin expression was already apparent in primary tumours (P = 0.030). The expression of vimentin significantly correlated with the grading of the primary tumour (P = 0.021). Conclusions: Our data indicate that the down‐regulation of E‐cadherin and the up‐regulation of nuclear β‐catenin expression might be crucial steps during tumour progression of tonsillar carcinomas, being already present in primary tumours in HPV‐driven carcinomas, but becoming apparent in HPV‐unrelated tumours later in the process of metastasis.     

Cyclin D1, E‐cadherin and beta‐catenin expression in FIGO Stage IA cervical squamous carcinoma: diagnostic value and evidence for epithelial–mesenchymal transition

Koay M H E, Crook M & Stewart C J R
(2012) Histopathology
Cyclin D1, E‐cadherin and beta‐catenin expression in FIGO Stage IA cervical squamous carcinoma: diagnostic value and evidence for epithelial–mesenchymal transition Aims: Immunohistochemistry is helpful in distinguishing cervical neoplastic lesions from their histological mimics, but has contributed less towards the sometimes problematic distinction of in‐situ and superficially invasive tumours. Epithelial–mesenchymal transition (EMT) may be a mechanism of invasion in cervical neoplasia and expression of EMT‐associated proteins could prove useful in this diagnostic setting. Methods and results: Immunohistochemical expression of cyclin D1, E‐cadherin and beta‐catenin was assessed in 22 biopsy specimens from FIGO Stage IA cervical squamous carcinomas, all of which also included foci of cervical intraepithelial neoplasia (CIN) 3, nine biopsies of CIN 3 adjacent to carcinoma, and 10 cases of CIN 3 only. Most invasive tumour cells expressed cyclin D1 and showed a reduction in E‐cadherin and beta‐catenin staining. Nuclear beta‐catenin expression was not observed. Cyclin D1 staining was reduced or showed altered distribution in most cases of CIN 3, while adhesion protein expression generally was preserved. However, altered protein expression similar to that of invasion was seen in some CIN lesions. Conclusions: Most superficially invasive cervical squamous carcinomas show immunophenotypical changes consistent with EMT. These alterations, particularly cyclin D1 expression, may be useful diagnostically. Similar changes in CIN 3 lesions may indicate the acquisition of increased invasive potential.     

Epithelial–mesenchymal transition and clinicopathological correlation in craniopharyngioma

Qi S‐T, Zhou J, Pan J, Zhang C, Silky C & Yan X‐R (2012) Histopathology 61 , 711–725
Epithelial–mesenchymal transition and clinicopathological correlation in craniopharyngioma Aims: To assess the immunophenotypic changes associated with epithelial–mesenchymal transition (EMT) in craniopharyngioma, especially at the tumour invasive front, and to correlate the findings with clinicopathological features and patient outcomes. Methods and results: Forty‐two craniopharyngiomas were investigated for the presence of EMT markers (vimentin, E‐cadherin and β‐catenin) by immunohistochemistry and western blot. The relationships between expression of these markers and various clinicopathological indicators and clinical outcomes of the tumours were analysed. There were statistically significant differences in the expression of vimentin and E‐cadherin–β‐catenin between adamantinomatous and papillary variants. The expression of vimentin and E‐cadherin (but not that of β‐catenin) in whole tumour sections was associated with tumour recurrence, and with postoperative weight and hypothalamic disturbances; the expression of vimentin and E‐cadherin–β‐catenin at the tumour invasive front was also associated with tumour recurrence, postoperative weight, and hypothalamic disturbances. The results from western blotting closely matched those of immunohistochemistry. Conclusions: Our study demonstrates, for the first time, the potential prognostic implications of vimentin, E‐cadherin and β‐catenin expression in craniopharyngiomas. EMT may represent a crucial mechanism in the progression of craniopharyngiomas.     

Expression of mismatch repair proteins, beta catenin, and E cadherin in intestinal-type sinonasal adenocarcinoma

BACKGROUND: Despite their histological resemblance to colorectal adenocarcinomas, there is little information about the molecular events involved in the pathogenesis of intestinal-type sinonasal adenocarcinomas (ITACs). AIMS: To evaluate the possible role of DNA mismatch repair (MMR) gene defects or disruptions of the E cadherin-beta catenin complex in ITAC by investigating the immunohistochemical expression of the MMR gene products, beta catenin, and E cadherin in a group of sporadic ITACs. METHODS: Ten sporadic cases of ITAC were stained with antibodies against MLH1, MSH2, MSH3, MSH6, beta catenin, and E cadherin. RESULTS: Nine cases showed strong nuclear expression of MLH1, whereas one case showed moderate staining. All 10 cases were strongly positive for MSH2 and MSH3. MSH6 was strong in nine cases, and moderate in one. Membranous beta catenin expression was strong in all 10 cases, and no case showed cytoplasmic or nuclear staining. E cadherin was strong in seven cases, and moderate in three cases. CONCLUSIONS: The preserved nuclear expression of MLH1, MSH2, MSH3, and MSH6 suggests that mutations or promoter methylation of MMR genes do not play a role in the pathogenesis of ITAC. The strong membranous staining for E cadherin and beta catenin and lack of abnormal cytoplasmic or nuclear expression is in keeping with the preservation of E cadherin-beta catenin complexes and beta catenin pathways.     

Loss of p53 expression is accompanied by upregulation of beta‐catenin in meningiomas: a concomitant reciprocal expression

Crosstalk between Wnt and p53 signalling pathways in cancer has long been suggested. Therefore in this study we have investigated the involvement of these pathways in meningiomas by analysing their main effector molecules, beta‐catenin and p53. Cellular expression of p53 and beta‐catenin proteins and genetic changes in TP53 were analysed by immunohistochemistry, PCR/RFLP and direct sequencing of TP53 exon 4. All the findings were analysed statistically. Our analysis showed that 47.5% of the 59 meningiomas demonstrated loss of expression of p53 protein. Moderate and strong p53 expression in the nuclei was observed in 8.5% and 6.8% of meningiomas respectively. Gross deletion of TP53 gene was observed in one meningioma, but nucleotide alterations were observed in 35.7% of meningiomas. In contrast, beta‐catenin, the main Wnt signalling molecule, was upregulated in 71.2%, while strong expression was observed in 28.8% of meningiomas. The concomitant expressions of p53 and beta‐catenin were investigated in the same patients. In the analysed meningiomas, the levels of the two proteins were significantly negatively correlated (P = 0.002). This indicates that meningiomas with lost p53 upregulate beta‐catenin and activate Wnt signalling. Besides showing the reciprocal relationship between proteins, we also showed that the expression of p53 was significantly (P = 0.021) associated with higher meningioma grades (II and III), while beta‐catenin upregulation was not associated with malignancy grades. Additionally, women exhibited significantly higher values of p53 loss when compared to males (P = 0.005). Our findings provide novel information about p53 involvement in meningeal brain tumours and reveal the complex relationship between Wnt and p53 signalling, they suggest an important role for beta‐catenin in these tumours.     

上皮型钙粘连素和环连蛋白在前列腺癌中的表达

目的　探讨上皮型钙粘连素 (E cadherin)和环连蛋白 (catenin)的表达与前列腺癌生物学行为及预后的关系。方法　以免疫组化法检测 6 7例人前列腺癌组织中E cadherin和catenin的表达。结果　前列腺癌E cadherin、α 、β 和γ catenin的低表达率分别为 4 0 3%、37 3%、6 0 %和 11 9% ,E cadherin和α catenin表达的减少与前列腺癌的病理分级、临床分期明显相关 ,E cadherin和α catenin低表达患者术后 5年生存率明显低于正常表达者。有淋巴结或骨转移的前列腺癌和无转移组的E cadherin和α catenin异常表达率差异显著。β catenin在前列腺癌组织中多为正常表达 ,仅有 4例呈细胞内低表达而核内表达却明显增加。 结论　E cadherin及α catenin有可能成为前列腺癌恶性程度及预后的评估指标 ,β catenin不仅参与细胞间的粘附作用而且在信号传导过程中具有重要意义。     

β‐Catenin and E‐cadherin expression in stage I adult‐type granulosa cell tumour of the ovary: correlation with tumour morphology and clinical outcome

Aims: To study E‐cadherin and β‐catenin expression in stage I adult‐type granulosa cell tumours (AGCTs) and correlate the findings with tumour morphology and clinical outcome. Methods and results: The study group comprised 62 FIGO stage I AGCTs, including 48 stage IA and 14 stage IC cases. Fifty patients (80.6%) had negative clinical follow‐up over periods from 3.0 to 19.2 years (median 6.4 years), and 12 patients (19.4%) developed metastases at intervals of 3.6–16.2 years (median 8.6 years). β‐Catenin and E‐cadherin were expressed in 62 (100%) and 53 (85%) primary tumours, respectively, and staining was more consistent and intense in areas showing sex cord‐like morphology. In contrast, diffuse tumour areas often showed weak or moderate staining (β‐catenin) or were negative (E‐cadherin), and there was reduced expression of both proteins in luteinized cells. Reduced β‐catenin expression in primary tumours correlated with increased risk of recurrence (P = 0.002) and a shorter time interval to recurrence, whereas there was no correlation between E‐cadherin staining and the risk of metastases. Conclusions: Localized variations in adhesion protein expression may partly explain the diverse morphological patterns exhibited by AGCT, and reduced β‐catenin staining in primary tumours may have value as an adverse prognostic factor.     

Correlation between invasive pattern and immunophenotypic alterations in endocervical adenocarcinoma

Stewart C J R, Crook M L, Little L & Louwen K
(2011) Histopathology 58 , 720–728
Correlation between invasive pattern and immunophenotypic alterations in endocervical adenocarcinoma Aims: To assess the immunophenotypic changes associated with epithelial‐mesenchymal transition (EMT) in endocervical adenocarcinoma, and correlate the findings with tumour morphology including growth pattern. Methods and results: Twenty‐seven endocervical adenocarcinomas were studied using a panel of immunohistochemical markers to vimentin, cyclin D1, E‐cadherin, beta‐catenin, p16 protein and cytokeratin 7. There were 24 moderately differentiated and three poorly differentiated tumours. Fourteen of the moderately differentiated carcinomas showed a focal infiltrative component, typically towards the deep tumour margin (invasive front), comprising attenuated glands, small cell clusters and single cells. These foci typically showed cytological alteration including loss of cellular polarity and cytoplasmic eosinophilia, while immunohistochemistry demonstrated reduced cell membrane E‐cadherin and beta catenin labelling, and expression of cyclin D1 and, in some cases, vimentin. Similar immunophenotypic changes were focally observed at the deep aspect of some larger ‘conventional’ tumour glands. No consistent changes were observed in the poorly differentiated carcinomas. Conclusions: Endocervical adenocarcinomas that demonstrate an infiltrative growth pattern show immunophenotypic changes consistent with EMT. Frequently, these are accompanied by a morphological alteration in the tumour cells and the changes exhibit a specific micro‐anatomical distribution. Epithelial‐mesenchymal transition may represent an important mechanism in the progression of some endocervical neoplasms.     

Inhibition of endocytosis blocks Wnt signalling to beta-catenin by promoting dishevelled degradation

Aim: The Wnt/Frizzled signalling pathway is highly conserved through evolution. Frizzled, the receptors for Wnts, have the topology of seven transmembrane spanning domain receptors. An important means of regulation of these receptors is internalization and desensitization through clathrin‐mediated endocytosis. Therefore, we investigated the effects of endocytosis inhibition on Frizzled4‐green fluorescent protein (FZD₄‐GFP) localization, dishevelled levels and Wnt‐3a signalling to β‐catenin. Methods: Experiments were performed in the mouse neuronal cell line SN4741 that has previously proven to be valuable for the investigation of Wnt/Frizzled signalling. FZD₄‐GFP distribution has been examined using confocal laser scanning microscopy. Dishevelled protein expression levels and the activation of β‐catenin upon treatment with endocytosis inhibitors (hyperosmolaric sucrose and K⁺ depletion), kinase inhibitors and Wnt‐3a were analysed by immunoblotting. Results: Hyperosmotic sucrose and K⁺ depletion increased the membrane localization of FZD₄‐GFP, and in parallel triggered fast (1–2 h) and almost complete (approx. 95%) degradation of endogenous dishevelled, which was independent of Wnt‐induced, CK1‐mediated phosphorylation of dishevelled. In addition, dishevelled depletion induced by endocytosis inhibition completely prevented canonical signalling by Wnt‐3a to β‐catenin even when osmotic conditions and endocytosis were reverted to normal. Conclusions: The data provide evidence for a molecular mechanism that could be a basis for a novel negative feedback loop within the Wnt/Frizzled pathway depending on dishevelled degradation. The identification of molecular details of regulatory mechanisms for the Wnt/Frizzled signalling pathway increases our understanding of pathway regulation, which might be of special physiological significance for embryonic development, cancer and neurological disorders.