Epithelial-mesenchymal Transition Phenomenon Research Articles

Molecular characterization of epithelial-mesenchymal transition and medical treatment related-genes in non-functioning pituitary neuroendocrine tumors.

Different medical therapies have been developed for pituitary adenomas. However, Non-Functioning Pituitary Neuroendocrine Tumors (NF-PitNET) have shown little response to them. Furthermore, epithelial-mesenchymal transition (EMT) has been linked to resistance to medical treatment in a significant number of tumors, including pituitary adenomas. We aimed to evaluate the expression of EMT-related markers in 72 NF-PitNET and 16 non-tumoral pituitaries. To further explore the potential usefulness of medical treatment for NF-PitNET we assessed the expression of somatostatin receptors and dopamine-associated genes. We found that SNAI1, SNAI2, Vimentin, KLK10, PEBP1, Ki-67 and SSTR2 were associated with invasive NF-PitNET. Furthermore, we found that the EMT phenomenon was more common in NF-PitNET than in GH-secreting pituitary tumors. Interestingly, PEBP1 was overexpressed in recurrent NF-PitNET, and could predict growth recurrence with 100% sensitivity but only 43% specificity. In parallel with previously reported studies, SSTR3 is highly expressed in our NF-PitNET cohort. However, SSTR3 expression is highly heterogeneous among the different histological variants of NF-PitNET with very low levels in silent corticotroph adenomas. NF-PitNET showed an enhanced EMT phenomenon. SSTR3 targeting could be a good therapeutic candidate in NF-PitNET except for silent corticotroph adenomas, which express very low levels of this receptor. In addition, PEBP1 could be an informative biomarker of tumor regrowth, useful for predictive medicine in NF-PitNET.

Stemness of breast cancer cells incubated on viscoelastic gel substrates

Acrylamide copolymer-based gel substrates with different viscoelasticity were employed to evaluate the viscoelasticity effect on the direct relation among cancer stemness and mesenchymal properties with induction of epithelial–mesenchymal transition (EMT) of human breast adenocarcinoma (MCF-7) cells in both normoxia and hypoxia. The softer gel substrate produced a large amount of surface molecule of cancer stem cells (CSC) marker CD44. In contrast, for the stem cell biomarker CD133 expression, their coefficient of damping (tan)-dependent manner was not contributed by EMT phenomenon and was an independent from acquisition of the EMT. The substrate damping as potential physical parameter emerged the important linkage to cancer stemness and EMT induction.

Role of RBMS3 Novel Potential Regulator of the EMT Phenomenon in Physiological and Pathological Processes.

RNA-binding protein 3 (RBMS3) plays a significant role in embryonic development and the pathogenesis of many diseases, especially cancer initiation and progression. The multiple roles of RBMS3 are conditioned by its numerous alternative expression products. It has been proven that the main form of RBMS3 influences the regulation of microRNA expression or stabilization. The absence of RBMS3 activates the Wnt/β-catenin pathway. The expression of c-Myc, another target of the Wnt/β-catenin pathway, is correlated with the RBMS3 expression. Numerous studies have focused solely on the interaction of RBMS3 with the epithelial–mesenchymal transition (EMT) protein machinery. EMT plays a vital role in cancer progression, in which RBMS3 is a new potential regulator. It is also significant that RBMS3 may act as a prognostic factor of overall survival (OS) in different types of cancer. This review presents the current state of knowledge about the role of RBMS3 in physiological and pathological processes, with particular emphasis on carcinogenesis. The molecular mechanisms underlying the role of RBMS3 are not fully understood; hence, a broader explanation and understanding is still needed.

Twist-related protein 1 induces epithelial-mesenchymal transition and renal fibrosis through the upregulation of complement 3.

We have demonstrated that complement 3 (C3) is upregulated and induces epithelial-mesenchymal transition (EMT) phenomenon and renal fibrosis in unilateral ureteral obstruction (UUO) kidney. We investigated roles of twist-related protein 1 (TWIST1) in EMT phenomenon and renal fibrosis through C3 upregulation in a mouse UUO model with gene silencer pyrrole-imidazole (PI) polyamides targeting TWIST1. We designed and synthesized PI polyamides targeting TWIST1 binding site on mouse pre-pro C3 promoter. Increased expression C3 mRNA with interferon-γ was significantly inhibited with PI polyamide in nephrotubular epithelial cells. Immunofluorescence showed suppression of E-cadherin and enhancement of α-smooth muscle actin (α-SMA) stainings as EMT phenomena in UUO kidney. TWIST1 and C3 expression was significantly increased in UUO kidney versus contralateral unobstructed kidney (CUK). Expression of transforming growth factor-β1 (TGF-β1), α-SMA and renin mRNAs was increased in UUO kidney versus CUK. Systemic administration of TWIST1 PI polyamide significantly suppressed increased C3 expression in UUO kidney versus CUK. PI polyamide administration also suppressed the increased expression of TGF-β1, α-SMA and renin mRNAs and histologically improved renal fibrosis in UUO kidney. These findings indicate that TWIST1 induces EMT phenomenon and renal fibrosis by TGF-β1 upregulation of C3 in mouse UUO model and that TWIST1 PI polyamide may be a novel medicine for renal fibrosis.

Adenosquamous and squamous cell carcinoma of the pancreas: two histopathological variants of ductal adenocarcinoma

Abstract Introduction: Primary squamous cell carcinoma (SCC) and adenosquamous carcinoma of the pancreas are rare malignancies for which diagnostic and treatment strategy are challenging. In this paper we present a literature review of these tumors based on two case reports. Case presentation: In the first case, a 55-year-old male presented with an inoperable pancreatic head/body junction tumor. Endoscopic ultrasound-guided fine needle aspiration was practiced, and histopathological examination revealed a squamous cell carcinoma of the pancreas. After exclusion of any another tumor, the diagnosis of cT4N0M0-staged primary pancreatic SCC was made. The patient is under treatment with gemcitabine and oxaliplatin. The second case is represented by a 73-year-old patient in which imagistic examinations highlighted a cystic mass of the pancreatic body. Following coporeo-caudal splenic-pancreatectomy and histopathological-proved diagnosis of adenosquamous carcinoma, the patient started chemotherapy but died at 11 months after surgery. Both tumor components displayed positivity for markers which prove ductal (cytokeratin19, maspin) and squamous differentiation (p63, cytokeratin5/6) same as vimentin, as indicator of epithelial mesenchymal transition (EMT). Conclusions: SCC and adenosquamous carcinoma of the pancreas are aggressive malignancies which prognosis remains highly reserved. These tumors might be variants of ductal adenocarcinomas which are dedifferentiated through EMT phenomenon.

Mechanism of SMND-309 Against Lung Injury Induced by Chronic Intermittent Hypoxia

Introduction: Obstructive sleep apnea-hypopnea syndrome (OSAHS) is a common sleep disorder that causes severe physiological disturbance. Lots of evidences showed that OSAHS was an important associated comorbidity that can affect pulmonary fibrosis patient’s survival. Until now, the potential mechanisms by which OSAS accelerate the progression of lung fibrosis are still unclear. Constructing a pathological model of chronic intermittent hypoxia (CIH), the present study aimed to explore the pathological progress and potential mechanism of lung injury caused by OSAHS. Meanwhile, SMND-309 was given for treatment to evaluate its potential therapeutic role in CIH induced lung injury. Methods: The mice were randomly divided into WT+RA group, WT+CIH group, SMND-309+RA group and SMND-309+CIH group. The WT+CIH group and SMND-309+CIH group were exposed in CIH condition for 12 weeks, while the other groups were processed in normal oxygen at the same time. SMND-309+RA group and SMND-309+CIH group were intraperitoneally injected with SMND-309 at the last week of modeling period. After 12 weeks of treatment, 3 mice of each group were perfused through the heart and lung tissues were isolated, fixed, sectioned and stained with HE, Masson and immunofluorescence stain. The rest of lung tissues were harvested for Western blot and Elisa assays. Results: The results showed that CIH treatment increased the expression of pro-inflammatory factor (TNF-α, IL-6), resulting in lung tissue structure disorder, inflammatory cell infiltration, increased pulmonary capillary permeability and pulmonary edema. The activation of NF-κB signaling pathway played a crucial role in the process of inflammation. Noticeably, we observed M2 macrophage accumulation in the lung after CIH exposure, which promoted the epithelial–mesenchymal transition (EMT) phenomenon and pulmonary tissue fibrosis process. Also, Elisa assays showed the increased expression of TGF-β, IL-10, IL-4 in CIH group. SMND-309 could inhibit pulmonary inflammation, reduce the accumulation of M2 macrophage, alleviate collagen deposition, and prevent the development of lung fibrosis. Conclusion: CIH could induce chronic lung inflammation, promote the activation of M2 macrophages, trigger the occurrence of EMT phenomenon and accelerate the deposition of lung collagen, eventually lead to lung tissue damage. This study presents a possible explanation by which interstitial lung diseases and particularly idiopathic pulmonary fibrosis (IPF) with OSAHS are usually associated with fast progress and poor prognosis. SMND-309 showed a good protective effect on CIH induced lung damage.

Pharmacokinetic evaluation and antitumor potency of liposomal nanoparticle encapsulated cisplatin targeted to CD24-positive cells in ovarian cancer.

CD24, which is upregulated in several human malignancies, is related to Epithelial-mesenchymal-transition (EMT) and has characteristics of cancer stem-like cells, especially in cisplatin-resistant ovarian carcinoma cells. Drug delivery systems represent a promising therapeutic approach for diseases with treatment resistance, and the present study investigated a novel CD24-targeted drug delivery system for advanced ovarian carcinoma. We produced liposomal cisplatin with a red fluorescent substance - cyanine 5.5 (GL-CDDP-Cy5.5). In order to target CD24-positive cells, an anti-CD24 monoclonal antibody was modified to the above drug (CD24-GL-CDDP-Cy5.5). Specific uptake of CD24-GL-CDDP-Cy5.5 was confirmed using a therapeutically resistant ovarian cancer cell line, Caov-3 cells. Antitumor effects of CD24-GL-CDDP-Cy5.5 were then evaluated in Caov-3 ×enograft mice. CD24-GL-CDDP-Cy5.5 showed more specific uptake by flow cytometry than GL-CDDP-Cy5.5. In xenograft mice, GL-CDDP-Cy5.5 and CD24-GL-CDDP-Cy5.5 treatment had significantly higher platinum concentration in disseminated tumor cells than cisplatin (P<0.05). Moreover, CD24-GL-CDDP-Cy5.5 suppressed tumor growth and prolonged survival time compared with other treatments. Median survival times of the control, cisplatin, GL-CDDP-Cy5.5 and CD24-GL-CDDP-Cy5.5 groups were 37, 36, 46 and 54 days after inoculation, respectively. Immunohistochemical analysis showed that CD24-GL-CDDP-Cy5.5 treatment, compared with GL-CDDP-Cy5.5, decreased the number of CD24-positive cells and suppressed the EMT phenomenon significantly (P<0.05). The present study demonstrated that CD24-GL-CDDP-Cy5.5, compared with other treatments, improved therapeutic efficacy. The present results suggested the potential for targeting anticancer therapeutics for CD24-positive cells to prevent disease progression.

Roles of Helicobacter pylori in the Epithelial-Mesenchymal Transition of Gastric Cancer

Gastric cancer (GC) is one of the most frequent malignant tumors in humans, with over 50% of patients after treatment suffering from recurrence and peritoneal metastasis. Helicobacter pylori (H. pylori) infection is critical to the development of GC. The phenomenon of epithelial-mesenchymal transition (EMT) in GC is linked with development of the invasive phenotype, which is very likely regulated by H. pylori through altering signaling pathways in the gastric cells. In this review, we conclude the current studies on how H. pylori affects the EMT of GC, thus contributing to its initiation and metastasis.

LMP1 Up-regulates Calreticulin to Induce Epithelial-mesenchymal Transition via TGF-β/Smad3/NRP1 Pathway in Nasopharyngeal Carcinoma Cells.

Background: Latent membrane protein 1 (LMP1) is known as an oncogenic protein encoded by the EBV genome. The purpose of this study was to investigate the mechanism of LMP1-induced cell epithelial-mesenchymal transition (EMT).Methods: The NP69 cell line of nasopharyngeal epithelial cells with high expression of LMP1 was established to observe the effect of high expression of LMP1 on cell growth, proliferation, cycle, apoptosis, migration and invasion. We used proteomics to screen and identify differentially expressed proteins related to LMP1-mediated epithelial cell transformation. Then, we analyzed the expression and significance of differentially expressed calreticulin (CRT) in nasopharyngeal carcinoma (NPC), and observed the effect of CRT expression on EMT in CNE2 cells of NPC. Finally, the expression of neuropilin-1 (NRP1), which is a protein downstream of the EMT-related signaling pathway TGF-β (transforming growth factor β), was detected.Results: LMP1 promoted NP69 cells proliferation, inhibited apoptosis and induced EMT. We identified 22 differentially expressed proteins associated with LMP1-induced EMT. Among them, CRT expression level was significantly increased in NPC compared with adjacent tissues, and was interrelated with TNM staging and lymph node metastasis of NPC. After knockdown of CRT expression, the phenomenon of cell EMT was reduced and the ability of cell migration and invasion was weakened. CRT regulated NRP1 expression by affecting SMAD3 phosphorylation.Conclusion: LMP1 induced cell EMT via TGF-β/Smad3/NRP1 pathway, which promoted migration and invasion of NPC cells.

Curcumin Inhibits the Migration and Invasion of Non-Small-Cell Lung Cancer Cells Through Radiation-Induced Suppression of Epithelial-Mesenchymal Transition and Soluble E-Cadherin Expression.

Radiotherapy has been reported to cause cancer metastasis. Thus, a new strategy for radiotherapy must be developed to avoid this side effect. A549 cells were exposed to radiation to induce an epithelial-mesenchymal transition (EMT) cell model. Real-time PCR and western blotting were used to detect mRNA and protein expression levels, and Transwell invasion and wound healing assays were used to detect cell migration and invasion. ELISA was used to detect soluble E-cadherin (sE-cad) secretion. siRNA was used to silence MMP9 expression. The results show that A549R cells exhibited an EMT phenotype with increased E-cadherin, N-cadherin, Snail, Slug, vimentin and Twist expression and decreased pan-keratin expression. sE-cad levels were increased in A549R cells and in the serum of NSCLC patients with distant metastasis. Exogenous sE-cad treatment and sE-cad overexpression promoted A549R and A549 cell migration and invasion. In contrast, blocking sE-cad attenuated A549 cell migration and invasion. Curcumin inhibited sE-cad expression and reversed EMT induced by radiation. Furthermore, curcumin suppressed sE-cad-enhanced A549 and A549R cell migration and invasion. Curcumin inhibited MMP9 expression, and silencing MMP9 suppressed sE-cad expression. Taken together, we found a nonclassic EMT phenomenon induced by radiation. Curcumin inhibits NSCLC migration and invasion by suppressing radiation-induced EMT and sE-cad expression by decreasing MMP9 expression.

Abstract 1035: PLEK2 promotes epithelial mesenchymal transition through increasing Snail1 in lung cancer cells

Abstract Pleckstrin 2 (PLEK2), one of the pleckstrin homology domain proteins, have been suggested to modulate actin rearrangement and cell spreading in T lymphocytes. Recently studies also implicated that PLEK2 paly an importance role in human myeloproliferative neoplasms, melanoma, and lung cancers. PLEK2 mRNA genes were related to poor overall survival in lung adenocarcinoma patients. However, the biological functions of PLEK2 in lung cancer is not clear. We detected the expression of PLEK2 in various lung adenocarcinoma cell lines using quantitative RT-PCR methods. Molecular manipulations were performed to investigate the roles of PLEK2 in lung cancer cells. Over-expression of PLEK2 in lung cancer cells exhibited markedly promoting in the proliferation and anchorage independent growth, whereas reduced of PLEK2 was found to suppress lung cancer cell progression. This studies also indicated that PLEK2 may play an important role in the epithelial-mesenchymal transition (EMT) features, including a morphological change, increased invasive and migratory ability, reduction the epithelial marker expression and up-regulated EMT regulators. Our results suggested that PLEK2 expression induce lung cancer cell EMT phenomenon through up-regulation Snail1 expression. These results will provide information and help us to understand the roles of PLEK2 in lung cancer cells. Citation Format: En-Chu Liu, Tsai-Tu Lee, Shey-Lin Wu, Tai-Lin Lee, Meng-Feng Tsai. PLEK2 promotes epithelial mesenchymal transition through increasing Snail1 in lung cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1035.

The cullin4A is up-regulated in chronic obstructive pulmonary disease patient and contributes to epithelial-mesenchymal transition in small airway epithelium

BackgroundChronic obstructive pulmonary disease (COPD) is a common respiratory disease with high morbidity and mortality. The most important pathophysiological change of COPD is airway obstruction. Airway obstruction can cause airflow restriction and obstructive ventilation dysfunction. Currently, many studies have shown that there is EMT phenomenon in the process of airway remodeling of COPD. Cullin4A (CUL4A) is an E3 ubiquitin ligase that interacts with other factors to form the E3 complex. Studies have shown that CLU4A is associated with EMT in non-small cell lung cancer and other cancers. However, its relationship with EMT in COPD has not been reported systematically. In this study, we detected the expression of CUL4A in lung epithelium of COPD patients. In addition, the regulatory effect and mechanism of CUL4A on EMT in COPD were clarified in small airway epithelial cells.MethodsThe expression of CUL4A was assessed by immunohistochemistry in lung epithelium specimens from smokers, non-smokers and patients with chronic obstructive pulmonary disease. The role of CUL4A on cigarette smoke extract (CSE)-induced epithelial-mesenchymal transition (EMT) in human small airway epithelial cells (HSAEpiCs) was assessed by silencing or overexpression CUL4A in vitro. Cigarette smoke is recognized as a high-risk factor in the induction of COPD, and its damage to the airway involves airway damage, airway inflammation and airway remodeling.ResultsThe results shown that CUL4A expression in small airway epithelium was significantly increased in patients with COPD. We also observed a significant negative association between CUL4A and FEV1%, a useful clinical marker for the diagnosis and evaluation of COPD severity, in small airway epithelial cells. In vitro, CSE-induced EMT is associated with high expression of CUL4A, and targeted silencing of CUL4A with shRNA inhibits CSE-induced EMT in human small airway epithelial cells.ConclusionsOur results showed that CUL4A was overexpressed in lung epithelium of COPD patients, and CUL4A could regulate EMT of human small airway epithelium, which revealed a new mechanism of remodeling of small airway epithelium of COPD patients.

Tumour budding : an emerging prognostic factor in colorectal carcinoma

Tumour budding, is a morphological feature seen in colorectal carcinoma and known to be associated with a poor outcome. It is thought to be representing the phenomenon of epithelial-mesenchymal transition of colorectal carcinoma. Colorectal malignancies exhibiting budding have shown an association with lymphatic invasion and lymph node metastasis. In addition, tumour budding has proven to be associated with tumour recurrence and is a negative prognostic indicator of overall survival of patients with colorectal carcinoma. Objectives of the study was to assess the prevalence of tumour budding in CRC (higher colorectal carcinoma) specimens and to determine the association between tumour budding and the histological grade, nodal involvement and tumour recurrence. A retrospective study was carried out at the Department of Pathology, Faculty of Medical Sciences, University of Sri Jayewardenepura. The study sample included 58 cases of colorectal carcinoma during the period January 2015 to June 2018. The histological grade, nodal status and tumour recurrence was recorded following review of histology slides. In addition, the slides were examined for the presence or absence of tumour budding, according to the ITBCC (November 2016) recommendations. Study population showed a tumour budding percentage of 32.75%. Tumour budding in colorectal carcinoma showed a statistically significant association with high histological grade, lymph nodal involvement and tumour recurrence (p-value Tumour budding in CRC is strongly predictive of high histological grade, lymph node metastases, and tumour recurrence. The study population included 02 cases of tumour recurrence in the budding group and a single case of cancer-related deaths.

Mechanism of epithelial-mesenchymal transition of pancreatic cancer induced by pancreatic stellate cell

Objective To explore the potential mechanism of epithelial-mesenchymal transition (EMT) of pancreatic cancer induced by pancreatic stellate cell (PSC). Methods The effect of PSC on EMT of pancreatic cancer was determined by direct co-culture. We also added signal pathway inhibitors and Notch-3 small interfere RNA (siRNA) to the co-culture system. Morphological change observation, two-chamber migration assays to assess the cell migration, fluorescent immunohistochemistry, Reverse transcriptase PCR (RT-PCR) and Western blotting to assess the expression level of Notch-3, E-cadherin, Vimentin was performed to determine whether the EMT happened. Results After co-culture with PSC, EMT phenomenon of PANC-1 happened with increased motility [mono-culture vs. co-culture, (16.4±2.6)/HP vs. (51.6±4.5)/HP, P=0.001]. L1790 (5 μmol/L, Notch signaling pathway inhibitor) can significantly inhibit the increased motility of PANC-1 cells, and suppress the occurrence of EMT. Notch-3 siRNA successfully inhibited Notch-3 gene expression, blocked the increased migration ability and EMT phenomenon of PANC-1 induced by PSC. Conclusion Notch-3 plays critical role in EMT of PANC-1 induced by PSC. Key words: Pancreatic cancer; Pancreatic stellate cell; Epithelial-mesenchymal transition

Abstract 492: Downregulation of microRNA-17-5p promotes EMT by targeting vimentin in colorectal cancer

Abstract Background: Colorectal cancer (CRC) is one of the most frequent forms of cancer worldwide. Cells with abnormal growth have the ability to invade or spread to other parts of the body (metastasis). Metastasis is the most common cause of death in CRC patients and activation of epithelial-mesenchymal transition (EMT) triggers metastasis. Thus, it is important to understand the mechanisms of EMT to increase the survival rate of CRC. MicroRNA-17 (miRNA-17) has been proven to be significantly higher in CRC tissues than normal tissues. However, there are few papers about the role of miRNA-17 in CRC metastasis. Therefore, in this study, we evaluated the mechanism underlying miRNA-17-5p related EMT in colon cancer cells. Methods: Expression level of miRNA-17-5p as well as target mRNA were analyzed in HCT-15, HT29, LoVo, SW480 and COLO205 cell lines. In order to evaluate the correlation between miRNA-17-5p and EMT phenomenon, selected cell lines were transiently transfected either miRNA-17-5p mimic or miRNA-17-5p inhibitor and then phenotyping studies were conducted. Moreover, to find out the direct target of miRNA-17-5p, we performed AGO2 immunoprecipitation in each colon cancer cell lines. Results: Real-time PCR revealed that miRNA-17-5p expression reversely correlated with vimentin expression in five colon cancer cell lines. Overexpression of miRNA-17-5p inhibited vimentin expression, cell migration and cell invasion in both LoVo and HT29 cells. On the other hand, inhibition of miRNA-17-5p promoted vimentin expression, cell migration and cell invasion in the same parallel cell lines. Through AGO2 immunoprecipitation analysis, we observed that miRNA-17-5p directly bind to vimentin 3'UTR to influence its stability and expression at the transcriptional level. Conclusion: Our data demonstrated that down-regulated miRNA-17-5p promotes colon cancer cell migration and invasion by regulating vimentin expression. These findings propose that miRNA-17-5p may serve as a potential prognostic biomarker in colorectal cancer. Citation Format: Tea Won Kim, Yeo Song Lee, Hye Kyung Hong, Su Jeong Song, Nak Hyeon Yun, Yong Beom Cho. Downregulation of microRNA-17-5p promotes EMT by targeting vimentin in colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 492.

Expression of β-catenin in regenerating renal tubules of cisplatin-induced kidney failure in rats.

β-Catenin is a multi-functional protein involved in nephrogenesis and also plays important roles in renal injury. Here, the expression of β-catenin was investigated in the proximal renal tubular epithelial cells in cisplatin (CDDP)-induced acute kidney injury (AKI) and chronic kidney injury (CKI), because CDDP-induced renal lesions were characterized by proximal renal tubular epithelial degeneration/regeneration and subsequent interstitial fibrosis. F344 rats were treated with CDDP. The expression of β-catenin and proliferative (Ki67) or fibrogenic [vimentin, α-smooth action (α-SMA)] markers was analyzed by immunolabeling. β-Catenin, vimentin and Ki67 were not seen in the proximal renal tubules of control rats. Interestingly, in CDDP-induced AKI, the regenerating proximal renal tubular epithelial cells reacting strongly with Ki67 expressed membranous or cytoplasmic β-catenin and also showed a positive reaction to vimentin but not to α-SMA. In CDDP-induced CKI, the epithelial cells of abnormally dilated or atrophied renal tubules did not react to β-catenin or Ki67, but showed positive reactions to vimentin and α-SMA. β-Catenin mRNAs were significantly increased in AKI and significantly decreased in CKI. Newly expressed β-catenin in the proximal renal tubules after AKI may participate in functional regeneration. In CKI, epithelial cells of abnormal renal tubules did not express β-catenin but reacted to vimentin, and α-SMA might indicate the epithelial-mesenchymal transition (EMT) formation, because α-SMA is usually expressed in myofibroblasts forming via EMT. The presence or absence of β-catenin expression would become a marker for the EMT phenomenon in progressive renal fibrosis.

Lipoxin A4 Suppresses Estrogen-Induced Epithelial-Mesenchymal Transition via ALXR-Dependent Manner in Endometriosis.

Epithelial-mesenchymal transition (EMT) is essential for embryogenesis, fibrosis, and tumor metastasis. Aberrant EMT phenomenon has been reported in endometriotic tissues of patients with endometriosis (EM). In this study, we further investigated the molecular mechanism of which lipoxin A4 (LXA4) suppresses estrogen (E2)-induced EMT in EM. The EMT markers were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot in eutopic endometrial epithelial cells (EECs) or investigated by immunohistochemistry and qRT-PCR in endometriotic lesion of EM mice. The invasion and migration under different treatments were assessed by transwell assays with or without Matrigel. The messenger RNA (mRNA) and activities of matrix metalloproteinase 2 (MMP-2) and MMP-9 were determined by qRT-PCR and gelatin zymography, respectively. Luciferase reporter assay was used to measure the activity of zinc finger E-box binding homeobox 1(ZEB1) promoter. The level of E2 in endometriotic tissues was assessed by enzyme-linked immunosorbent assay. In eutopic EECs, stimulatory effects of E2 on EMT progress, migration, and invasion were all diminished by LXA4. Lipoxin A4 reduced E2-induced ZEB1 promoter activity. Lipoxin A4 also attenuated the phosphorylation of extracellular signal-regulated kinase and p38 mitogen-activated protein kinase induced by E2. Co-incubation with Boc-2 rather than DMF antagonized the influence of LXA4. Animal experiments showed that LXA4 inhibited the EMT progress, MMP expression, and proteinase activities of endometriotic lesion in an LXA4 receptor (ALXR) manner, which suppressed the progression of EM. ZEB1 mRNA expression was upregulated and well correlated with E2 level in human endometrium. Lipoxin A4 suppresses E2-induced EMT via ALXR-dependent manner in eutopic EECs, which reveals a novel biological effect of LXA4 in EM.

Tumor-associated macrophages correlate with phenomenon of epithelial-mesenchymal transition and contribute to poor prognosis in triple-negative breast cancer patients

BackgroundTumor-associated macrophages (TAMs) are associated with poor outcomes in multiple solid cancers and play important roles in cancer progression. Epithelial-mesenchymal transition (EMT) may account for metastasis and recurrence. However, the association between TAMs and EMT is not clarified in triple-negative breast cancer (TNBC). The aim of this study was to investigate the effects of TAMs on EMT in TNBC. Material and methodsWe studied specimens from 278 patients with TNBC. TAMs marker cluster of differentiation 163 and EMT-related marker E-cadherin were detected by immunohistochemistry in TNBC tissues, and their clinical significance was evaluated from the patients' medical records. ResultsTNBC patients with polarized cluster of differentiation 163+ TAMs infiltration and low level of E-cadherin had a significantly higher risk of aggressive features, including recurrence, histologic differentiation, and lymph node metastasis. Infiltration of TAMs was also negatively correlated with E-cadherin in TNBC tissues. Multivariate analysis indicated that infiltration of TAMs and low expression of E-cadherin were independent prognostic factors of overall survival and disease-free survival in TNBC patients. ConclusionsHigh infiltration of TAMs was associated with low expression of E-cadherin and could be used as an unfavorable prognostic factor for patients with TNBC.

The cellular character of liquefaction degeneration in oral lichen planus and the role of interferon gamma

Oral lichen planus (OLP) is a T-cell-mediated chronic inflammatory oral mucosal disease of unknown etiology, and liquefaction degeneration in the basal keratinocytes is one of the specific histological changes. However, the understanding of liquefaction degeneration is still very limited, and how does it affect the prognosis of LP is largely unknown. Therefore, the objective of this study was to clarify the intrinsic change behind the liquefaction degeneration in lichen planus and to evaluate the effect of the OLP-typical cytokine, IFN-γ, on these changes. Biopsies were collected from patients with OLP; immunochemistry staining was performed to analyze E-cadherin, vimentin, CK19, β1 integrin, nestin, STAT1, and STAT3 expression. Primary oral epithelial cells were cultured in vitro, and 20 ng/mL IFN-γ was applied to assay the effect on epithelial cells. E-cadherin expression was decreased but vimentin expression was increased in the OLP epithelial cells that undergo liquefaction degeneration, showing the typical epithelial-mesenchymal transition (EMT) alteration. In vitro research showed that the OLP-typical cytokine, IFN-γ, possesses EMT-inducing ability, and the primary oral epithelial cells stimulated by IFN-γ acquired some properties of cancer stem cells, expressing more β1 integrin, α6 integrin, and nestin. In addition, the major downstream mediator of IFN-γ receptor, STAT1, was expressed more intensive and extensive with the malignant transition of OLP. Liquefaction degeneration in oral lichen planus is an EMT phenomenon, the IFN-γ may be the main inducer, and IFN-γ signaling might be implicated in malignant transition of OLP.

Abstract 911: Destabilizing β-catenin and Ras could be an ideal therapy for treating metastatic colorectal cancer

Abstract INTRODUCTION Aberrant activations of Wnt/β-catenin and Ras signaling pathways by APC (90%) and K-Ras (40-50%) mutations are closely interacted and accelerate CRC tumorigenesis. Protein level of β-catenin as well as Ras is increased by APC loss via inhibiting GSK3B-mediated poly-ubiquitination dependent Ras degradation through recruitment of β-TrCP E3 linker. Moreover, additional K-Ras mutation results in the liver metastasis by secondary activation of the Wnt/β-catenin signaling pathways via Ras-ERK and -AKT pathways in addition to the initial activation by APC loss. Consequently, both β-catenin and Ras were highly increased in human CRC patient tissues, especially in metastatic tumor. EXPERIMENT PROCEDURES To investigate the synergistic effects of aberrant Wnt/β-catenin and Ras signaling pathways on tumor development to metastasis, we established the APCMin/+/K-RasG12DLA2 compound mouse that harbor APC and K-Ras mutations in animal model. We also analyzed Ras protein level using human CRC patient tissues of normal mucosa and adenocarcinoma (N=24) and metastatic adenocarcinoma (N=26) and tumor budding (N=10). To recapitulate the metastatic tumor environment which have enriched receptor tyrosine kinases (RTKs), we maintained and experimented with EGF (20ng/ml) in every in vitro studies. RESULTS We observed that both β-catenin and Ras were increased in adenocarcinoma and metastatic adenocarcinoma of human CRC compared with normal mucosa. Interestingly, both β-catenin and Ras were most significantly increased in tumor budding regions which histopathologically represent epithelial mesenchymal transition (EMT). In murine model, the EMT phenomenon was effectively induced and invasion of small intestinal tumors were occurred in APCMin/+/K-RasG12DLA2 with increased level of β-catenin and Ras compared with APCMin/+. However, EMT phenomenon and its following events such as migration and invasion of CRC cells induced by APC and K-Ras mutations were abolished by destabilization of β-catenin and Ras. CONCLUSION Our results show that destabilizing β-catenin and Ras could be effective therapeutic targets for inhibiting metastasis in the early stage of CRC development Citation Format: Yong-Hee Cho, Jeong-Soo Yoon, Kang-Yell Choi. Destabilizing β-catenin and Ras could be an ideal therapy for treating metastatic colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 911. doi:10.1158/1538-7445.AM2017-911