Granulocyte colony‐stimulating factor‐producing pancreatic anaplastic carcinoma in ascitic fluid at initial diagnosis: A case report

Granulocyte colony‐stimulating factor (G‐CSF)‐producing pancreatic tumors are extremely rare. These tumors have an aggressive clinical course and no established treatment. Here, we report an autopsy case of G‐CSF‐production in pancreatic anaplastic carcinoma (PAC). A 72‐year‐old woman presented with a large pancreatic head mass and multiple liver metastases. Laboratory data showed leukocytosis (leukocyte count 113.3 × 10³/µL) and high serum G‐CSF levels (441 pg/mL; normal range: <39.0 pg/mL). The ascitic fluid was submitted to our pathology laboratory at initial diagnosis. Cytopathology showed that smears from the ascitic fluid were highly cellular and contained numerous malignant cells, mainly in loose groupings. Occasional pseudoglandular formations and giant cells were also present. The malignant cells were round, and no spindle‐shaped cells were visible. The nuclei were round to ovoid with coarsely granular chromatin and large prominent nucleoli. Upon immunocytochemistry, tumor cells were positive for G‐CSF and vimentin; there was no E‐cadherin expression. Histopathological examination of the tumor showed a mixed composition of adenocarcinomatous and sarcomatous regions. Upon immunohistochemistry, both components were positive for G‐CSF. Few CD34‐positive myeloblasts were observed in the bone marrow. Thus, we diagnosed this as a case of G‐CSF production in PAC with leukocytosis. To the best of our knowledge, this is the first report on G‐CSF expression immunocytochemically confirmed in PAC. Diagn. Cytopathol. 2017;45:463–467. © 2017 Wiley Periodicals, Inc.     

Mesenchymal–epithelial transition of pancreatic cancer cells at perineural invasion sites is induced by Schwann cells

Epithelial–mesenchymal transition (EMT) promotes invasion and metastasis of pancreatic ductal adenocarcinoma (PDAC). However, the importance of its reverse process, mesenchymal–epithelial transition (MET), for PDAC remains unclear. We aimed to characterize the histological finding “focal differentiation” in PDAC at perineural invasion sites in the context of MET and to investigate the role of Schwann cells in inducing tumor MET. Tumor differentiation and immunohistochemical expressions of E‐cadherin, SMAD3, and vimentin at perineural invasion sites were examined in 168 PDAC tissues. Four PDAC cell lines were co‐cultured with Schwann cells to investigate cell morphology, motility, or EMT‐related markers using immunocytochemistry and quantitative PCR. Of 168 tumors, 124 (74%) showed focal differentiation with enhanced E‐cadherin membrane expression (P < 0.001) and decreased nuclear accumulation of SMAD3 (P < 0.001). Among 115 PDACs harboring grade 1/2 tumor, tumors with focal differentiation showed worse survival compared to those without focal differentiation (P = 0.019). PDAC cells co‐cultured with Schwann cells demonstrated a sheet‐like appearance, increased E‐cadherin expression, decreased expressions of SMAD3 and vimentin, and reduced cell motility. In conclusion, MET‐like change is induced by Schwann cells, suggesting that Schwann cells contribute to PDAC colonization in pancreatic nerves through activating the MET machinery inside tumor cells in the pancreatic tumor microenvironment.     

An epithelial to mesenchymal transition programme does not usually drive the phenotype of invasive lobular carcinomas

Epithelial to mesenchymal transition (EMT) is a cellular phenotype switching phenomenon which occurs during normal development and is proposed to promote tumour cell invasive capabilities during tumour progression. Invasive lobular carcinoma (ILC) is a histological special type of breast cancer with a peculiar aetiology – the tumour cells display an invasive growth pattern, with detached, single cells or single files of cells, and a canonical feature is the loss of E‐cadherin expression. These characteristics are indicative of an EMT or at the very least that they represent some plasticity between phenotypes. While some gene expression profiling data support this view, the tumour cells remain epithelial and limited immunohistochemistry data suggest that EMT markers may not feature prominently in ILC. We assessed the expression of a panel of EMT markers (fibronectin, vimentin, N‐cadherin, smooth muscle actin, osteonectin, Snail, Twist) in 148 ILCs and performed a meta‐analysis of publically available molecular data from 154 ILCs. Three out of 148 (2%) ILCs demonstrated an early and coordinated alteration of multiple EMT markers (down‐regulation of E‐cadherin, nuclear TWIST, and up‐regulation of vimentin, osteonectin, and smooth muscle actin). However, the data overall do not support a role for EMT in defining the phenotypic peculiarities of the majority of ILCs. Copyright © 2015 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.     

Immunohistochemical analysis of the expression of E‐cadherin and ZEB1 in non‐small cell lung cancer

We performed an immunohistochemical analysis of the expression of zinc‐finger E‐box binding homeobox 1 (ZEB1), a master regulator of epithelial‐mesenchymal transition (EMT), and determined its relationship with E‐cadherin in 157 non‐small cell lung carcinomas (93 adenocarcinomas, 36 squamous cell carcinomas, 18 large cell carcinomas, and 10 pleomorphic carcinomas). Although the expression of E‐cadherin was low in the subset of adenocarcinomas (10%) and squamous cell carcinomas (11%), ZEB1 expression was only observed in one case of squamous cell carcinoma and none of the adenocarcinomas. In contrast, the low expression of E‐cadherin (50% and 90%, respectively) and the positive expression of ZEB1 (11% and 50%, respectively) were more frequently observed in poorly differentiated carcinomas (large cell carcinomas and pleomorphic carcinomas). Overall, the expression of ZEB1 was inversely correlated with that of E‐cadherin. Furthermore, the distribution of ZEB1‐positive cancer cells was more restricted than in the area in which the expression of E‐cadherin was lost, and the former was detected within the latter. We concluded that the expression of ZEB1 was not necessarily associated with the low expression of E‐cadherin in lung adenocarcinomas and squamous cell carcinomas. The expression of ZEB1 correlated with an undifferentiated and/or sarcomatoid morphology that may occur in the late stage of EMT.     

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.     

Slug regulates E‐cadherin expression in metastatic adenocarcinoma cells isolated from pleural fluid

Slug protein is a key regulator of epithelial‐mesenchymal transition, but its expression in cancer is less well studied. To evaluate the expression of slug, E‐cadherin and vimentin in adenocarcinoma cells from malignant pleural effusions, we analyzed 121 malignant pleural fluid specimens. Twenty‐eight nonmalignant pleural fluid specimens were analyzed as control. Besides clinical cytological diagnosis tests, immunofluorescence, immunocytochemistry and Western blotting methods were used. Results showed strong membrane staining of E‐cadherin in adenocarcinoma cells from pleural fluid. Slug mainly showed nucleus staining. Cytoplasma positive of vimentin was found in adenocarcinoma cells isolated from pleural fluid. Slug, E‐cadherin and vimentin expression was found in 43/121 (36%), 87/121 (72%) and 102/121 (84%) cases, respectively. Our data showed elevated levels of slug were accompanied by down regulation of E‐cadherin and the expression of vimentin in adenocarcinoma cells. In addition, there was no relationship between slug expression and patient's age or gender or tumor site. Hyperplasia epithelium cells from nonmalignant pleural fluid were uniformly negative for E‐cadherin and slug. In conclusion, the results demonstrated the inverse expression of slug and E‐cadherin in the majority of malignant pleural fluid cases compared with nonmalignant pleural fluid. The slug protein may be helpful to access the prognosis of patients with pleural fluid. Diagn. Cytopathol. 2013. © 2011 Wiley Periodicals, Inc.     

Epithelial–mesenchymal transition markers in malignant ovarian germ cell tumors

The purpose of this study was to determine the expression and potential clinical role of epithelial‐to‐mesenchymal transition (EMT)‐related factors in malignant ovarian germ cell tumors (MOGCT). Protein expression of E‐cadherin, N‐cadherin, P‐cadherin, Zeb1, HMGA2, and vimentin by immunohistochemistry was analyzed in 42 MOGCT from patients treated in Norway during the period 1981–2001. Expression was analyzed for association with clinicopathologic parameters. E‐cadherin (p = 0.016) and HMGA2 (p = 0.002) expression was significantly higher in immature teratomas and yolk sac tumors compared with dysgerminomas. Vimentin (p < 0.001) and Zeb1 (p = 0.029) staining was significantly higher in immature teratomas compared with yolk sac tumors and dysgerminomas, whereas no significant differences were observed for N‐cadherin and P‐cadherin. EMT‐associated markers were not significantly related to clinicopathologic parameters including age, tumor diameter, and FIGO stage. In conclusion, based on this limited series, EMT‐associated markers are not associated with clinical parameters in MOGCT, in contrast to ovarian carcinoma. EMT‐related proteins are differentially expressed among various MOGCT subtypes, suggesting differences in biological characteristics associated with invasion and metastasis.     

Thymidylate synthase is functionally associated with ZEB1 and contributes to the epithelial‐to‐mesenchymal transition of cancer cells

Thymidylate synthase (TS) is a fundamental enzyme of nucleotide metabolism and one of the oldest anti‐cancer targets. Beginning from the analysis of gene array data from the NCI‐60 panel of cancer cell lines, we identified a significant correlation at both gene and protein level between TS and the markers of epithelial‐to‐mesenchymal transition (EMT), a developmental process that allows cancer cells to acquire features of aggressiveness, like motility and chemoresistance. TS levels were found to be significantly augmented in mesenchymal‐like compared to epithelial‐like cancer cells, to be regulated by EMT induction, and to negatively correlate with micro‐RNAs (miRNAs) usually expressed in epithelial‐like cells and known to actively suppress EMT. Transfection of EMT‐suppressing miRNAs reduced TS levels, and a specific role for miR‐375 in targeting the TS 3'‐untranslated region was identified. A particularly relevant association was found between TS and the powerful EMT driver ZEB1, the shRNA‐mediated knockdown of which up‐regulated miR‐375 and reduced TS cellular levels. The TS–ZEB1 association was confirmed in clinical specimens from lung tumours and in a genetic mouse model of pancreatic cancer with ZEB1 deletion. Interestingly, TS itself appeared to have a regulatory role in EMT in cancer cells, as TS knockdown could directly reduce the EMT phenotype, the migratory ability of cells, the expression of stem‐like markers, and chemoresistance. Taken together, these data indicate that the TS enzyme is functionally linked with EMT and cancer differentiation, with several potential translational implications. Copyright © 2017 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.     

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.     

A unique pancreatic ductal adenocarcinoma with carcinosarcomatous histology, immunohistochemical distribution of hCG-beta, and the elevation of serum alpha-feto-protein

Pancreatic undifferentiated carcinomas with a neoplastic mesenchymal component (carcinosarcoma) have not been well described to date. The author experienced an autopsy case of a unique pancreatic ductal adenocarcinoma with carcinosarcomatous histology. The patient was a 90 year old Japanese male who died of cahexia with generalized tumor extension. Post-mortem examinations revealed some distinctive or representative components discerned in the tumor tissue. One was the well differentiated ductal adenocarcinoma. The second and the major finding was undifferentiated short spindle shaped or small round sarcomatous cells, which lacked an epithelial nature but showed positivity for CD10+, CD56+, Ki67++, p53++, and were focally positive for Desmin and vimentin. These two components were mixed and constituted the histology of the carcinosarcoma. In another area, anaplastic, large, pleomorphic tumor cells showed the focal immunohistochemical distribution of alpha-feto-protein and human chorionic gonadotropin. An ultrastructural study revealed adenocarcinoma cells with apical mucin secreting granules and well developed ductal differentiation, whereas undifferentiated sarcomatous cells showed primitive fibroblastic or mesenchymal characters without specific differentiation. Conclusively these findings suggested that this well differentiated adenocarcinoma gradually enlarged, accumulated genetic alternations, and then transformed into large and undifferentiated tumor cells, rapidly growing small sarcomatous cells, and a histology of carcinosarcoma.     

Fine-needle aspiration cytology of carcinosarcoma of the parotid gland: Cytohistological and immunohistochemical findings

Carcinosarcoma or true malignant mixed tumor of salivary glands is a very rare neoplasm which shows malignant features of the epithelial and mesenchymal components. We hereby present cytological, pathological, and immunohistochemical findings of one such case, first examined by fine-needle aspiration cytology. The aspirate was cell-rich and consisted of large cell clusters which at low magnification showed an arrangement reminiscent of a jigsaw puzzle. An amorphous substance was observed in the middle of some of these clusters. There were also numerous dissociated cells which often displayed marked atypia. Histology showed a tumor with malignant epithelial and mesenchymal components. the carcinomatous areas consisted of cells arranged in solid nests or glandular structures. the sarcomatous areas showed osteosarcoma, low-grade chondrosarcoma, and undifferentiated components. On immunohistochemistry, the tumor coexpressed epithelial and mesenchymal markers. Cytokeratin, keratin, and epithelial membrane antigen were mainly localized to the carcinomatous portion, whereas vimentin and neuron-specific enolase were restricted to the sarcomatous areas. S-100 protein was positive in both. Our findings support previous views that this tumor may be related to pleomorphic adenoma. © 1995 Willey- Liss, Inc.     

miR-183 promotes radioresistance of lung adenocarcinoma H1299 cells via epithelial-mesenchymal transition

Lung adenocarcinomas are usually sensitive to radiation therapy, but some develop resistance. Radiation resistance can lead to poor patient prognosis. Studies have shown that lung adenocarcinoma cells (H1299 cells) can develop radioresistance through epithelial-mesenchymal transition (EMT), and this process is regulated by miRNAs. However, it is unclear which miRNAs are involved in the process of EMT. In our present study, we found that miR-183 expression was increased in a radioresistant lung adenocarcinoma cell line (H1299R cells). We then explored the regulatory mechanism of miR-183 and found that it may be involved in the regulation of zinc finger E-box-binding homeobox 1 (ZEB1) expression and mediate EMT in lung adenocarcinoma cells. qPCR results showed that miR-183, ZEB1, and vimentin were highly expressed in H1299R cells, whereas no difference was observed in E-cadherin expression. Western blot results showed that ZEB1 and vimentin were highly expressed in H1299R cells, while E-cadherin expression was decreased. When miR-183 expression was inhibited in H1299R cells, radiation resistance, proliferation, and cell migration were decreased. The expression of ZEB1 and vimentin in H1299R cells was decreased, while the expression of E-cadherin was increased. Moreover, miR-183 overexpression in H1299 cells enhanced radiation resistance, proliferative capacity, and cell migration ability. The expression of ZEB1 and vimentin in H1299 cells was increased, while that of E-cadherin was decreased. In conclusion, miR-183 may promote EMT and radioresistance in H1299 cells, and targeting the miR-183-ZEB1 signaling pathway may be a promising approach for lung cancer treatment.     

Liquid‐based sputum cytology of bicomponent mucin‐producing adenocarcinoma of the trachea with histologic comparison

Primary tracheal adenocarcinomas are rare. Here, we report on the liquid‐based sputum cytology of a bicomponent mucin‐producing tracheal adenocarcinoma with histologic and immunohistochemical comparison. A 72‐year‐old man presented with dyspnea. Computed tomography revealed a lobulated mass in the mid‐trachea. Sputum cytology showed clusters of atypical cuboidal cells that have pleomorphic and hyperchromatic nuclei and intracytoplasmic mucin. There were additional bland‐looking components of regular clusters of cuboidal epithelial cells. These cells were initially evaluated as benign cells. Sleeve resection revealed a 3.7‐cm‐sized transmural mass, composed of tubulopapillary cuboidal epithelial cell structures. Half of the tumor comprised of bland‐looking epithelial cells with small nuclei and eosinophilic cytoplasm; the remainder comprised of hyperchromatic cells with larger, distinct nucleoli. Direct interfaces between these components were observed, and both of these components equally displayed the characteristics of invasion and p53 expression. Two cell types in sputum cytology were retrospectively evaluated as adenocarcinomas of different grades. The patient has not shown recurrence for 8 months, postoperatively. Diagn. Cytopathol. 2016;44:1120–1124. © 2016 Wiley Periodicals, Inc.