Expression Of Epithelial-mesenchymal Transition-related Proteins Research Articles

Fibroblast growth factor 3 promotes spontaneous mammary tumorigenesis in Tientsin albino 2 mice via the FGF3/FGFR1/STAT3 pathway.

Tientsin albino 2 (TA2) mice can develop spontaneous breast cancer (SBC), which is associated with multiple pregnancies and infection with the mouse mammary tumor virus (MMTV). In this study, we sought to elucidate the molecular mechanisms underlying the development of SBC in TA2 mice induced by MMTV. The integration site of MMTV in TA2 SBC was identified using whole-genome sequencing. The expression of fibroblast growth factor 3 (FGF3) in SBCs and normal breast tissues was compared. The primary cell line, TA-1106, derived from SBC, was cultured. The proliferation, cell cycle, migration, invasion, and tumorigenicity abilities, as well as the expression of epithelial-mesenchymal transition-related proteins, phosphorylated STAT3, and phosphorylated Akt, were assessed in MA-891cell line from TA2 and TA-1106 cells after FGF3 knockdown. The binding of FGF3 to FGF receptor 1 (FGFR1) was determined by co-immunoprecipitation. Additionally, the relationship between STAT3 and Akt phosphorylation was investigated using a small molecule inhibitor and STAT3 knockdown. MMTV integrated upstream of the FGF3 gene, and the FGF3 protein was highly expressed in TA2 SBCs. FGF3 knockdown in MA-891 and TA-1106 decreased their proliferation, migration, and invasion abilities, affected the cell cycle and expression of epithelial-mesenchymal transition-related proteins, and inhibited the growth of animal xenografts. FGF3 binds to FGFR1, and either FGF3 or FGFR1 knockdown decreases STAT3 and Akt phosphorylation levels. Inhibition of phosphorylation or expression of STAT3 resulted in decreased Akt phosphorylation levels. Inhibition of Akt phosphorylation also resulted in decreased STAT3 phosphorylation levels. Furthermore, treatment of MA-891 and TA-1106 cells with Wortmannin or Stattic caused FGFR1 upregulation in addition to inhibiting Akt or STAT3 phosphorylation. The results of this study demonstrate that FGF3 plays a significant role in the development of SBC through the FGF3/FGFR1/STAT3 signaling pathway. There is a reciprocal activation between STAT3 and Akt. Inhibition of STAT3 or Akt phosphorylation promoted the expression of FGFR1. Validating the conclusions obtained in this study in human breast cancer (HBC) may contribute to targeted therapy and it is worth exploring whether the homologous sequences of MMTV in HBC have a similar oncogenic effect.

MP04-16 EPITHELIAL TO MESENCHYMAL TRANSITION PROMOTES PD-L1 EXPRESSION IN RENAL CELL CARCINOMA

You have accessJournal of UrologyCME1 Apr 2023MP04-16 EPITHELIAL TO MESENCHYMAL TRANSITION PROMOTES PD-L1 EXPRESSION IN RENAL CELL CARCINOMA Allison May, Jonathan Jiang, Tyler Robinson, and Evan Keller Allison MayAllison May More articles by this author , Jonathan JiangJonathan Jiang More articles by this author , Tyler RobinsonTyler Robinson More articles by this author , and Evan KellerEvan Keller More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003215.16AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Sarcomatoid dedifferentiation in renal cell carcinoma (RCC) is thought to occur when the parental cell type, such as clear cell RCC (ccRCC), undergoes an epithelial mesenchymal transition (EMT). The mesenchymal sarcomatoid cells have increased ability to invade and metastasize, and thus sarcomatoid RCC (sRCC) has a very poor prognosis. Interestingly however, there is encouraging evidence that sRCC responds at higher rates to PD-1/PD-L1 directed immunotherapy than ccRCC. We aim to test the hypothesis that EMT directly upregulates PD-L1 expression in sRCC and that PD-L1 may have positive feedback on induction of EMT. METHODS: Baseline expression of EMT markers, EMT transcription factors and PD-L1 were measured using Western Blot and qPCR in two clear cell RCC lines (Caki-1 and 786-O) and five sarcomatoid RCC lines (RCJ-41T1, RCJ-41T2, RCJ-41M, UOK-276, UOK-127). EMT was induced in the cells through treatment with multiple agents including TGFB, IFNg, HGF, SPP1, and CoCl2 and by over expression of Snail or Zeb1. Effect of EMT on PD-L1 expression was measured. To evaluate the role of PD-L1 on EMT, PD-L1 expression was decreased using siRNA and its effect on EMT related protein expression was measured using Western Blot. RESULTS: At baseline, sRCC cell lines generally had higher expression of EMT related markers and transcription factors than ccRCC lines. Baseline PD-L1 expression positively correlated with increasing EMT state in all cell lines. Treatment of cells with known EMT inducers, TGFB, IFNg, and hypoxia led to increased levels of mesenchymal marker N-cadherin, decreased epithelial marker E-cadherin, and increased levels of EMT transcription factors Zeb1, Snail, and Slug. Treatment of cells with HGF and SPP1, two molecules associated with sRCC based on our prior work, also induced EMT. Finally, over expression of known EMT transcription factors Snail and Zeb1 led to EMT as well. All methods of EMT induction concomitantly increased PD-L1 expression. Knockdown of PD-L1 was tested in UOK-276 and 786-O cells and led to decreased expression levels of EMT transcription factors and increased epithelial markers. CONCLUSIONS: These data support the theory that sRCC arises from EMT and that EMT directly leads to upregulation of PD-L1. Furthermore, these results suggest that PD-L1 maintains the mesenchymal state. These findings may help explain the enhanced response to PD-1/PD-L1 directed therapy in sRCC and suggest that markers of EMT state could represent a biomarker for immunotherapy response in RCC. Source of Funding: Clarke Family Fellowship for Kidney Cancer Research © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e39 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Allison May More articles by this author Jonathan Jiang More articles by this author Tyler Robinson More articles by this author Evan Keller More articles by this author Expand All Advertisement PDF downloadLoading ...

Characteristics and clinical significance of polyploid giant cancer cells in laryngeal carcinoma.

ObjectivesWe aimed to construct an induction system for polyploid giant cancer cells (PGCCs), as well as to investigate PGCC features and clinical significance.MethodsA laryngeal neoplasm‐PGCC induction system was constructed using paclitaxel liposomes (PTX). We used western blots to compare expression of epithelial‐mesenchymal transition‐related proteins, stem cell interrelated proteins, and cyclin‐associated proteins. We then measured PGCC count in tissue samples of patients with laryngeal neoplasms and analyzed its relationship with prognosis. Statistical significance was determined using t‐tests.ResultsPTX successfully induced PGCCs. Western blotting showed that CyclinB1, CDC25C, CDK1, E‐cadherin, and EIF‐4A expression decreased in PGCCs compared with normal cancer cells, whereas vimentin and CD133 expression increased. Number of PGCCs in laryngeal cancer tissues and overall survival time were inversely correlated (P < .05).ConclusionsPTX successfully induces PGCC formation in laryngeal carcinoma, which may be the cause of poor prognosis in patients with laryngeal cancer.Level of Evidence: 4.

Effect of MicroRNA-210 on the Growth of Ovarian Cancer Cells and the Efficacy of Radiotherapy

Objective: The objective of this study is to explore the role of miR-210 in the growth of ovarian cancer cells and the correlation with radiotherapy and to elucidate underlying molecular mechanisms. Methods: Human ovarian cancer cell lines OVCAR3 and SKOV3 were cultured in vitro, and miR-210 over-expression and low-expression ovarian cancer cell models were established by cell transfection. MTT assay was used to detect the proliferation activity. Transwell was used to detect the migration and invasion abilities. Western blot measured the expression of proteins related to cell proliferation, migration, and invasion. The cells were treated with different doses of ionizing radiation, and then the cell proliferation activity was detected by MTT. The expression of apoptosis-related proteins was detected by Western blot. The Caspase-Glo^® Kit was used to detect the activity of cellular caspase 3/7 enzymes. Results: The proliferation, migration, and invasion abilities of miR-210 over-expression ovarian cancer cells were increased (p < 0.05), the expressions of PTEN and E-cadherin were decreased, and the expression of p-Protein kinase B (AKT), N-cadherin, Snail, and Vimentin were elevated. After ionizing radiation, the sensitivity of miR-210 over-expression cells to radiotherapy was decreased, the expression of apoptosis-related protein Bax was decreased, the expression of Bcl-2 was increased, and the activity of cellular caspase 3/7 enzyme was reduced (p < 0.05). Conclusion: miR-210 can promote the proliferation, migration, and invasion of ovarian cancer cells by activating the AKT signaling pathway and regulating the expression of Epithelial-mesenchymal transition-related proteins. miR-210 can reduce the sensitivity of ovarian cancer cells to radiotherapy by inhibiting apoptosis, which might serve as a potential target for the treatment of ovarian tumors.

Silencing SIX1 inhibits epithelial mesenchymal transition through regulating TGF-β/Smad2/3 signaling pathway in papillary thyroid carcinoma

ObjectiveTo investigate the sineoculis homeobox homolog 1 (SIX1) affect the epithelial mesenchymal transition (EMT) in papillary thyroid carcinoma (PTC) through regulating TGF-β/Smad2/3 signaling pathway. MethodsThe SIX1 expression in cytological specimens, tissues or PTC cell lines was detected by qRT-PCR, western blotting or immunohistochemistry. A series of vitro experiments including flow cytometry, CCK-8, wound-healing and Transwell were used to evaluate the biological characteristics in a PTC cell line (NPA cells), which were divided into Blank, Negative control (NC), SIX1, SIX1-siRNA, LY-364947 (TGF-β/Smad2/3 pathway inhibitor) and SIX1 + LY-364947 groups. TGF-β/Smad2/3 pathway and EMT related protein expression were measured by qRT-PCR and western blotting. ResultsSIX1 mRNA expression was increased in cytological specimens from PTC patients as compared with the non-toxic nodular goitre (NTG) patients. Moreover, compared with adjacent normal tissues, expressions of SIX1, N-cadherin and Vimentin were higher while E-cadherin was lower in PTC tissues; and SIX1 was positively correlated with N-cadherin and Vimentin but was negatively correlated with E-cadherin. Furthermore, the SIX1 expression was associated with histopathology, extrathyroidal extension (ETE), lymph node metastasis (LNM), pT stage, TNM stage, and distant metastasis. In addition, the expressions of TGFβ1, p-SMAD2/3, N-cadherin and Vimentin were downregulated in NPA cells after LY-364947 treatment with upregulated E-cadherin, decreased cell proliferation and metastasis, and enhanced cell apoptosis, which was reversed by SIX1 overexpression. ConclusionSilencing SIX1 can inhibit TGF-β/Smad2/3 pathway, thereby suppressing EMT in PTC, which may be a novel avenue for the treatment of PTC.

Correlation of JMJD3 and Epithelial-mesenchymal Transition-related Protein Expression in Breast Cancer and Clinical Significance

Correlation of JMJD3 and Epithelial-mesenchymal Transition-related Protein Expression in Breast Cancer and Clinical Significance

Role of platelet infiltration as independent prognostic marker for gastric adenocarcinomas.

BackgroundIn recent years, the relationship between malignant tumors and platelets has been paid more attention. The increase of platelets is an independent risk factor for the poor prognosis of some malignant tumors.MethodsThis study retrospectively analyzed the clinical and pathological data of 114 patients with initial gastric cancer from August 2005 to August 2018 in Shandong Provincial Hospital. Single‐factor and multifactor survival analysis were used to evaluate the effect of platelet elevation on postoperative survival. The gastric cancer tissues of the Jinan Central Hospital and its matched paracancerous tissues were collected. The expression of platelets in tissues was detected by immunofluorescence technique. Different numbers of platelets were co‐cultured with MKN‐45 cells, CCK‐8 assay and transwell assay were performed, and the expression of epithelial‐mesenchymal transition‐related proteins was detected.ResultsPlatelet count was independent factors affecting prognosis. The stratified analysis showed that there was a statistically significant difference in the 5‐year survival rate between the platelet‐increase group and the normal platelet group in the TNM stages I‐II. The expression of platelets in gastric cancer tissues was higher than that in adjacent tissues. The results of CCK‐8 and transwell showed that platelets significantly enhanced the proliferation and metastasis capability of MKN‐45 cells in a concentration‐dependent manner. After co‐culture, the expression level of E‐cadherin protein in MKN‐45 cells decreased, and the protein expression levels of N‐cadherin, vimentin, and VEGFA increased.ConclusionPlatelet elevation is closely related to the occurrence, development, and metastasis of gastric cancer, and platelet count can be used as a prognostic indicator for malignant tumors.

Scavenger receptor class A, member 5 is associated with thyroid cancer cell lines progression via epithelial-mesenchymal transition.

Thyroid cancer (TC) has become one of most common endocrine malignancies in recent decades. Due to gene background polymorphism, it's outcome goes quite differently in each patient. For exploring the mechanism, we performed whole transcriptome sequencing of paired papillary thyroid carcinoma (PTC) and adjacent thyroid tissues. As a result, scavenger receptor class A member 5 (SCARA5) might be a crucial anti‐oncogene associated with PTC. By RT‐qPCR, we first detected the expression of SCARA5 in PTC tissue and three type of TC cell lines. Besides, The Cancer Genome Atlas (TCGA) data were gathered to analysis the relationship between SCARA5 and clinical feature. A series of loss‐function experiments in TC cell lines (KTC‐1 and BCPAP) to investigate the function of SCARA5 in PTC. The results showed that SCARA5 expression in PTC was lower than adjacent normal tissue. And, it's consistent with the TCGA database. After analyse the correlation between SCARA5 expression and clinicopathological features in TCGA database, we discovered that downregulated SCARA5 is significantly connected age (P = .04) and tumour size (P = .032). Knockdown of SCARA5 in TC cell line could significantly increase the function of cells proliferation, colony formation, migration, and invasion. Furthermore, we also proved that SCARA5 could modulate the expression of epithelial‐mesenchymal transition‐related proteins, which influence invasion and migration. To best of our knowledge, SCARA5 is a suppressor gene which was associated with PTC and might be a potential therapeutic target in the future.Significance of the studyThyroid cancer (TC) has become one of most common endocrine malignancies in recent decades. By whole transcriptome sequencing of paired papillary thyroid carcinoma (PTC) and adjacent thyroid tissues, author discovered that scavenger receptor class A member 5 (SCARA5) might be crucial anti‐oncogene associated with PTC. Furthermore, knocking‐down of SCARA5 in TC cell line can increase the function of cells proliferation, colony formation, migration, and invasion. Author also proved that SCARA5 could modulate the expression of epithelial‐mesenchymal transition‐related proteins.

Dehydroandrographolide Inhibits Osteosarcoma Cell Growth and Metastasis by Targeting SATB2-mediated EMT.

The 12-hydroxy-14-dehydroandrographolide (DP) is a predominant component of the traditional herbal medicine Andrographis paniculata (Burm. f.) Nees (Acanthaceae). Recent studies have shown that DP exhibits potent anti-cancer effects against oral and colon cancer cells. This investigation examined the potential effects of DP against osteosarcoma cell. A cell analyzer was used to measure cell viability. The cell growth and proliferation were performed by Flow cytometry and BrdU incorporation assay. The cell migration and invasion were determined by wound healing and transwell assay. The expression of EMT related proteins was examined by Western blot analysis. In this study, we found that DP treatment repressed osteosarcoma (OS) cell growth in a dose-dependent manner. DP treatment significantly inhibited OS cell proliferation by arresting the cell cycle at G2/M phase. In addition, DP treatment effectively inhibited the migration and invasion abilities of OS cells through wound healing and Transwell tests. Mechanistic studies revealed that DP treatment effectively rescued the epithelialmesenchymal transition (EMT), while forced expression of SATB2 in OS cells markedly reversed the pharmacological effect of DP on EMT. Our data demonstrated that DP repressed OS cell growth through inhibition of proliferation and cell cycle arrest; DP also inhibited metastatic capability of OS cells through a reversal of EMT by targeting SATB2. These findings demonstrate DP's potential as a therapeutic drug for OS treatment.

Growth‐associated protein 43 promotes thyroid cancer cell lines progression via epithelial‐mesenchymal transition

Thyroid cancer is maintaining at a high incidence level and its carcinogenesis is mainly affected by a complex gene interaction. By analysis of the next‐generation resequencing of paired papillary thyroid cancer (PTC) and adjacent thyroid tissues, we found that Growth Associated Protein 43 (GAP43), a phosphoprotein activated by protein kinase C, might be novel markers associated with PTC. However, its function in thyroid carcinoma has been poorly understood. We discovered that GAP43 was significantly overexpressed in thyroid carcinoma and these results were consistent with that in The Cancer Genome Atlas (TCGA) cohort. In addition, some clinicopathological features of GAP43 in TCGA database showed that up‐regulated GAP43 is significantly connected to lymph node metastasis (P < 0.001) and tumour size (P = 0.038). In vitro experiments, loss of function experiments was performed to investigate GAP43 in PTC cell lines (TPC‐1 and BCPAP). The results proved that GAP43 knockdown in PTC cell significantly decreased the function of cell proliferation, colony formation, migration, and invasion and induced cell apoptosis. Furthermore, we also indicated that GAP43 could modulate the expression of epithelial‐mesenchymal transition‐related proteins, which could influence invasion and migration. Put those results together, GAP43 is a gene which was associated with PTC and might be a potential therapeutic target.

Antitumor effect of kurarinone and underlying mechanism in small cell lung carcinoma cells.

BackgroundKurarinone, a prenylated flavonone isolated from the roots of Sophora flavescens, is known to be cytotoxic against many human cancer cells but not human small cell lung carcinoma (SCLC) yet. Also, the exact molecular mechanism of kurarinone for induction cytotoxicity remains unknown.Material and methodsWe investigated the effects of kurarinone on cell proliferation, apoptosis, and migration in H1688 SCLC cells. Cell viability was determined by the MTT assay. Apoptotic indices such as cell cycle, mitochondrial membrane potential, cytochrome c release, caspase activity, and death receptors were evaluated by flow cytometry. Transwell migration and invasion assays were also included.ResultsOur results indicated that kurarinone significantly decreased H1688 cell viability and induced the accumulation of sub-G1 fractions by activating caspase-3, -9, and PARP cleavage accompanied by the elevated release of cytochrome c and mitochondrial dysfunction in H1688 cells. Additionally, kurarinone promoted Fas and TRAIL receptor-1 and -2 expression via the caspase-8/Bid pathway, suggesting that kurarinone triggered apoptosis via the mitochondria-mediated and receptor-mediated apoptotic pathways. We also observed that kurarinone repressed migration and invasion capabilities of SCLC cells by suppressing the expression of epithelial-mesenchymal transition-related proteins and matrix metalloproteinases.ConclusionOur findings provided evidence that kurarinone can induce apoptosis in SCLC cells via multiple mechanisms and delayed the cell migration and invasion of SCLC cells.

NECTIN4 promotes papillary thyroid cancer cell proliferation, migration, and invasion and triggers EMT by activating AKT.

Papillary thyroid cancer (PTC) is the most frequent type of malignant thyroid cancer, but its molecular mechanisms remain unknown. To better understand the tumorigenesis and progression of PTC, we conducted a comprehensive analysis of the whole-transcriptome resequencing of paired PTC and normal thyroid tissues. Nectin cell adhesion molecule 4 (NECTIN4) was significantly overexpressed in thyroid carcinoma compared with that in matched normal tissue. We also assessed the relation between the expression level of NECTIN4 and the clinicopathological features of PTC in The Cancer Genome Atlas database, and results showed that upregulated NECTIN4 is associated with lymph node metastasis (P<0.001) and tumor size (P=0.017). The biological function of NECTIN4 was also investigated by using the PTC cell lines TPC-1 and KTC-1. In vitro experiments demonstrated that NECTIN4 downregulation significantly inhibits the colony formation, proliferation, migration, and invasion of PTC cell lines. NECTIN4 could modulate the expression of epithelial–mesenchymal transition-related proteins via the PI3K/AKT pathway, and SC79, an AKT phosphorylation activator, could reverse the si-RNA knockdown effect. In addition, after the use of AKT inhibitors (LY 294,002), we found that SiRNA have similar effect with AKT inhibitors. Taking the results together, the current study shows that NECTIN4 has important biological implications in the tumorigenesis and metastasis of PTC and may be a potential therapeutic target for the disease.

LncRNA ADPGK-AS1 promotes pancreatic cancer progression through activating ZEB1-mediated epithelial–mesenchymal transition

ABSTRACTObjective: This study was conducted to investigate the effects of ADP dependent glucokinase antisense RNA 1 (ADPGK-AS1)/ miR-205-5p/ zinc finger E-box binding homeobox 1 (ZEB1) on PC cells.Methods: Differentially expressed lncRNAs and miRNAs in pancreatic cancer (PC) were identified by microarray analysis. In silico ceRNA analysis was conducted to find out the interactions among lncRNAs, miRNAs and mRNAs. Quantitative real-time PCR (qRT-PCR) was utilized to examine the expression of miR-205-5p and lncRNA ADPGK-AS1 in PC and non-cancerous cells. The association between miR-205-5p and ADPGK-AS1 as well as miR-205-5p and ZEB1 was determined by dual-luciferase reporter gene assay. After manipulating the expression of ADPGK-AS1, mir-205-5p and ZEB1 in PANC-1 and SW-1990 cells, cell proliferation, migration, invasion and apoptosis were respectively confirmed by cell counting kit-8 (CCK-8) assay, transwell assay and TUNEL. Western blot was applied to examine the expression of Epithelial-mesenchymal Transition-related proteins. In vivo experiment was conducted to further determine the effect of miR-205-5p/ZEB1 on tumorigenic ability of PC cells.Results: MiR-205-5p was low-expressed while ZEB1 and ADPGK-AS1 were high-expressed in PC tissues and cells compared with the normal. Dual-luciferase reporter gene assay proved that ADPGK-AS1 could directly target miR-205-5p and miR-205-5p could directly target ZEB1 3′UTR. The expression of MiR-205-5p was negatively correlated with proliferation, migration and invasion, and positively correlated with apoptosis rate of PC cells, while ZEB1 and ADPGK-AS1 had an inversed effect. Further in vitro and in vivo investigation indicated that epithelial-mesenchymal transition (EMT) could be restrained by miR-205-5p through targeting ZEB1. ADPGK-AS1 strongly promoted the tumorigenesis via downregulating miR-205-5p expression and induced the EMT process in vivo.Conclusion: ADPGK-AS1 inhibited miR-205-5p and therefore promoted PC progression through activating ZEB1-induced EMT.

Comparison of EMT mediated tyrosine kinase inhibitor resistance in NSCLC

In the United States, lung cancer is the second most common cancer in men and women. In 2017, 222,500 new cases and 155,870 deaths from lung cancer are estimated to have occurred. A tyrosine kinase receptor, epidermal growth factor receptor (EGFR), is over expressed or mutated in non-small cell lung cancer (NSCLC) resulting in increased cell proliferation and survival. Tyrosine kinase inhibitors (TKIs) are currently being used as therapy for NSCLC patients, however, they have limited efficacy in NSCLC patients due to acquisition of resistance. This study investigates the role of epithelial-mesenchymal transition (EMT) in the development of resistance against TKIs in NSCLC. Currently, the role of p120-catenin, Kaiso factor and PRMT-1 in reversal of EMT in T790M mutated and TKI-resistant NSCLC cells is a new line of study. In this investigation we found upregulation of cytoplasmic p120-catenin, which was co-localized with Kaiso factor. In the nucleus, binding of p120-catenin to Kaiso factor initiates transcription by activating EMT-transcription factors such as Snail, Slug, Twist, and ZEB1. PRMT-1 was also found to be upregulated, which induces methylation of Twist and repression of E-cadherin activity, thus promoting EMT. We confirmed that TKI-resistant cells have mesenchymal cell type characteristics based on their cell morphology and gene or protein expression of EMT related proteins. EMT proteins, Vimentin and N-cadherin, displayed increased expression, whereas E-cadherin expression was downregulated. Finally, we found that the knockdown of p120-catenin and PRMT-1 by siRNA or use of a PRMT-1 inhibitor Furamidine increased Erlotinib sensitivity and could reverse EMT to overcome TKI resistance.

Study on Attenuating Angiogenesis and Epithelial-Mesenchymal Transition (EMT) of Non-Small Cell Lung Carcinoma (NSCLC) by Regulating MAGEC2.

Objective:To investigate the role of MAGE family member C2 in angiogenesis and epithelial–mesenchymal transition of non-small cell lung carcinoma.Methods:The Cancer Genome Atlas data set was analyzed to filter the highly expressed gene melanoma antigen family C2 in non-small cell lung carcinoma. Quantitative reverse transcription-polymerase chain reaction was performed to verify the overexpression of melanoma antigen family C2 in non-small cell lung carcinoma cell lines. Melanoma antigen family C2 complementary DNA and short hairpin RNA (shRNA) were transfected into SK-MES-1 cells to regulate melanoma antigen family C2 expression. Cell Counting Kit-8 assay, flow cytometry, wound healing assay, and Transwell assay were performed to investigate the effect of melanoma antigen family C2 on proliferation, apoptosis, migration, and invasion of SK-MES-1 cell line. Western blot was used to detect the expression of epithelial–mesenchymal transition markers. Enzyme-linked immunosorbent assay was performed to investigate the secretion of vascular endothelial growth factor, and tube formation assay was conducted to explore the effect of melanoma antigen family C2 on angiogenesis ability of the tumor. Tumor xenograft on nude mice and immunohistochemical/hematoxylin and eosin staining were also performed to detect the influence of melanoma antigen family C2 on growth and metastasis of non-small cell lung carcinoma cells.Results:Melanoma antigen family C2 was highly expressed in non-small cell lung carcinoma cells; melanoma antigen family C2 promoted the expression of epithelial–mesenchymal transition-related proteins as well as enhance the secretion of vascular endothelial growth factor and promote angiogenesis; melanoma antigen family C2 promoted proliferation, migration, and invasion and suppressed apoptosis of non-small cell lung carcinoma cells. It could also facilitate growth and metastasis of non-small cell lung carcinoma in vivo.Conclusion:Melanoma antigen family C2 was a critical factor of angiogenesis and epithelial–mesenchymal transition in non-small cell lung carcinoma.

Long Non-Coding RNA MALAT1 Decreases the Sensitivity of Resistant Glioblastoma Cell Lines to Temozolomide

Background/Aim: Multidrug resistance (MDR) is largely responsible for the failure of chemotherapy. The long non-coding RNA (lncRNA) metastasis-associated lung adenocarcinoma transcript (MALAT1) has been reported to be closely related to tumor biology. In the present study, whether MALAT1 contributes to the resistance of glioblastoma cell lines to temozolomide (TMZ) was investigated. Methods: The glioblastoma cell lines U251 and U87 were exposed to increasing concentrations of TMZ to generate TMZ-resistant colonies (the U251/TMZ and U87/TMZ cell lines). The expression levels of MALAT1 and proteins related to epithelial-mesenchymal transition (EMT) were detected by real-time PCR and western blot, respectively. After the transfection of si-MALAT1 or pcDNA-MALAT1, cell viability, mRNA expression of MDR-associated proteins (MDR1, MRP5 and LRP1), and protein expression of EMT related proteins (ZEB1, Snail and SLUG) were evaluated. Results: The expression of MALAT1 was upregulated in the U251/TMZ and U87/TMZ cell lines compared to that in U251 and U87 cell lines, respectively. The treatment of si-MALAT1 decreased MDR1, MRP5, and LRP1 expression, enhanced cell sensitivity to TMZ, and downregulated ZEB1 protein expression, whereas pcDNA-MALAT1 had the opposite effects. However, the effects of si-MALAT1 on MDR -associated protein expression, cell viability, and EMT status were reversed by the transfection of pcDNA-ZEB1, and the effects of pcDNA-MALAT1 were reversed by the transfection of si-ZEB1. In vivo, MALAT1 overexpression enhanced tumors’ TMZ resistance and upregulated ZEB1 expression. Conclusion: MALAT1 decreased the sensitivity of resistant glioma cell lines to TMZ by regulating ZEB1.

Correlation of overexpression of nestin with expression of epithelial-mesenchymal transition-related proteins in gastric adenocarcinoma.

Nestin is associated with neoplastic transformation. However, the mechanisms by which nestin contributes regarding invasion and malignancy of gastric adenocarcinoma (GAC) remain unknown. Recent studies have shown that the epithelial-mesenchymal transition (EMT) is important in invasion and migration of cancer cells. In the present study, we aimed to investigate the expression of nestin and its correlation with EMT-related proteins in GAC. The expression of nestin and EMT-related proteins was examined in GAC specimens and cell lines by immunohistochemistry and Western blotting. Clinicopathological features and survival outcomes were retrospectively analyzed. Positive nestin immunostaining was most obviously detected in the cytoplasm, nucleus or both cytoplasm and nucleus of tumor cells in 19.2% (24/125) of GAC tissues, which was significantly higher than that in normal gastric mucosa tissues (1.7%, 1/60) (p=0.001). Nestin expression was closely related to several clinicopathological factors and EMT-related proteins (E-cadherin, vimentin and Snail) and displayed a poor prognosis. Interestingly, simultaneous cytoplasmic and nuclear nestin expression correlated with EMT-related proteins (E-cadherin, vimentin and Snail) (p<0.05) and lymph node metastasis (p=0.041) and a shorter survival time (p<0.05), but this was not the case with cytoplasmic or nuclear nestin expression. Nestin, particularly expression in both cytoplasm and nucleus, might be involved in regulating EMT and malignant progression in GAC, with potential as an unfavorable indicator in tumor diagnosis and a target for clinical therapy.

Abstract LB-186: Twist-1 overexpression modulates cancer progression according to the microsatellite instability status in colorectal cancer cell lines

Abstract Background: The Epithelial-mesenchymal transition (EMT) is a process by which epithelial cells lose their cell polarity and cell-cell adhesion, and gain migratory and invasive properties to become mesenchymal cells. EMT has also been shown to occur in the initiation of metastasis for cancer progression. Twist-1 has driving effects in cancer metastasis and is involved in the process of EMT. Overexpression of Twist-1 is common in metastatic carcinomas. Moreover, Twist-1 plays an important role in some physiological processes involved in metastasis, like tumor cell migration, angiogenesis, invasion, chromosomal instability and maintenance of cancer stem cells via the induction of EMT. In this study, we evaluated if Twist-1 and its downstream signaling cascades induced activation of EMT according to the microsatellite instability status in colon cancer cell lines. Methods: To understand this mechanism, LS513 (MSS) and LoVo (MSI) cells were transfected with Twist-1-pCMV plasmids and GFP-pCMV plasmid was used as negative control. Western blots were used to detect expression of EMT related proteins. Cell proliferation assay, cell migration assay, cell invasion assay, and colony forming assay were used to confirm the metastatic effect by Twist-1 overexpression. Also, we examined tumorigenesis assay in vivo to evaluate cancer growth induced by Twist-1 overexpression. Results: The expression of E-cadherin was down-regulated, but that of vimentin was up-regulated in both Twist overexpressed-LS513 and LoVo cell lines. There were mesenchymal morphological changes such as spindle like shape in both cell lines. We found that overexpression of Twist-1 induced downregulation of E-cadherin and upregulation of β-catenin via AKT phosphorylation in these cells. Also, we identified the upregulation of NF-kB as a mediator of the cancer progression in Twist-1 overexpressed LS513 cells. However, the expression of NF-kB decreased in Twist-1 overexpressed LoVo cell lines. Cell proliferation, migration, invasion and in vivo tumorigenecity were increased more in Twist-1 activated cells than negative control cells in LS513 cell line. However, there were no differences of colony forming assay and in vivo tumor growth between Twist-1 activated cells and negative control cells in LoVo cell line although cell proliferation, cell migration, and cell invasion were increased in Twist-1 activated cell lines. Conclusion: Twist-1 overexpression could induce activation of EMT in colon cancer cell lines. NF-kB played as a mediator of cancer progression in Twist-1 overexpressed LS513 (MSS) cells. But, NF-kB expression was down-regulated in Twist-1 overexpressed-LoVo (MSI) cells. These results suggest that upregulation of Twist-1 is associated with colon cancer progression via activation of NF-kB signaling pathway in MSS colon cancers but not in MSI colon cancers. Citation Format: Sungwon Jung, Hye Kyung Hong, Bo Young Oh, So Young Kim, Heunmin Song, Heewon Park, Woo Yong Lee, Yong Beom Cho. Twist-1 overexpression modulates cancer progression according to the microsatellite instability status in colorectal cancer cell lines. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr LB-186. doi:10.1158/1538-7445.AM2014-LB-186

MP31-16 LAMININ BINDING GLYCAN DEPLETION ON α-DYSTROGLYCAN IN PROSTATE CANCER CELLS PROMOTES EPITHELIAL-MESENCHYMAL TRANSITION AND ENHANCES TUMOR FORMATION

You have accessJournal of UrologyProstate Cancer: Basic Research II1 Apr 2014MP31-16 LAMININ BINDING GLYCAN DEPLETION ON α-DYSTROGLYCAN IN PROSTATE CANCER CELLS PROMOTES EPITHELIAL-MESENCHYMAL TRANSITION AND ENHANCES TUMOR FORMATION Tohru Yoneyama, Shingo Hatakeyama, Yuki Tobisawa, Takuya Koie, Chikara Ohyama, and Minoru Fukuda Tohru YoneyamaTohru Yoneyama More articles by this author , Shingo HatakeyamaShingo Hatakeyama More articles by this author , Yuki TobisawaYuki Tobisawa More articles by this author , Takuya KoieTakuya Koie More articles by this author , Chikara OhyamaChikara Ohyama More articles by this author , and Minoru FukudaMinoru Fukuda More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2014.02.925AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES α-dystroglycan (α-DG) represents a highly glycosylated cell surface molecule that is expressed in the epithelial cell-basement membrane (BM) interface and plays an essential role in epithelial cell-BM interaction. α-DG mediated epithelial cell-BM interaction is often impaired in aggressive prostate cancer (PCa), yet roles and underlying mechanisms in PCa progression remain unclear. We report here a suppressor function of laminin binding (LB) glycan on α-DG in PCa progression. METHODS PC3 cells express varying amounts of LB glycans attached to α-DG. After 2 consecutive cell sorting, PC3 cells were separated into LB glycan expressing substantial (PC3-H) and minimal (PC3-L) amount of LB glycans. Epithelial-mesenchymal transition (EMT) related gene expression profiles of PC3-H and PC3-L were analyzed by Affymetrix GeneChip human Gene1.0 ST Array. Expression of EMT related proteins were also analyzed immunoblotting. To evaluate LB glycan function in PCa progression, we performed cell motility, matrigel invasion assay and orthotopic tumor formation assay. RESULTS We found that LB glycan depletion caused cell shape change. PC3-L has a mesenchymal shape (spindle-like) and downregulated E-cadherin while upregulated N-cadherin and vimentin. PC3-H cells reverted to an epithelial shape (rounded) and expressed epithelial cell markers. Matrigel invasion assay showed PC3-L cells invaded much faster than PC3-H cells. Strikingly, PC3-L cells produced much bigger tumor after being inoculated in the orthotopic prostate of SCID mice and much metastasis to the draining lymph nodes compared with PC3-H and parental PC3 cells. ERK/AKT phosphorylation is also increased PC3-L tumor compare with PC3-H. CONCLUSIONS These results indicate that the LB glycan depletion on prostate cancer cells promotes EMT and correlated to the progression of PCa. © 2014FiguresReferencesRelatedDetails Volume 191Issue 4SApril 2014Page: e328 Peer Review Report Advertisement Copyright & Permissions© 2014MetricsAuthor Information Tohru Yoneyama More articles by this author Shingo Hatakeyama More articles by this author Yuki Tobisawa More articles by this author Takuya Koie More articles by this author Chikara Ohyama More articles by this author Minoru Fukuda More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Desmoplastic Melanoma

Desmoplastic melanoma (DM) is a rare variant of melanoma. Most frequently, it seems as clinically ambiguous and histologically characterized by a poorly demarcated neoplasm composed of a proliferation of spindle melanocytes dispersed in a prominent collagenous stroma. It often represents a diagnostic challenge, delaying its detection. We analyzed the expression profile of 29 (28 "pure" and 1 "combined") DM. These data were compared with a series of 62 primary vertical growth phase nondesmoplastic melanomas (NDMs) using a set of proteins including melanocytic markers (S-100 protein and melan-A) and epithelial-mesenchymal transition (EMT)-related proteins (E-cadherin, N-cadherin, SPARC, WT1, and PKCα). The S-100 protein confirmed the melanocytic origin of the DM (positive in 96%). The significant positive expression of N-cadherin, SPARC, and WT1 in DM (61%, 82%, and 71%) compared with NDM (28%, 43%, and 47%; P < 0.05) and a lower expression of E-cadherin in DM (14%) compared with NDM (61%) support specific adhesive and migratory properties of DM tumor cells. The study was carried out with tissue microarrays that partly limited the study of the tumor sections. This study demonstrates, for the first time, a prominent expression of epithelial-mesenchymal transition-related proteins in DMs and tries to be one more step in refining its knowledge and leading to a better understanding of its biological and clinical behaviors.