scholarly journals Spontaneous malignant transformation of ovarian surface epithelial cells correlates with EMT alteration and stemness acquisition

2019 ◽  
Author(s):  
Bingyan Li ◽  
Hemei Zhang ◽  
Yi Guo ◽  
Lizhi Liu ◽  
Yongfeng Hou ◽  
...  
Keyword(s):  
Epithelial Mesenchymal Transition ◽  
Cell Model ◽  
Mrna Level ◽  
Neoplastic Transformation ◽  
Surface Epithelium ◽  
Ovarian Carcinomas ◽  
Self Renewal ◽  
Mesenchymal Transition ◽  
Ovarian Surface

Abstract Background In order to complete the physiological function of postovulatory repair during repeated ovulation, the ovarian surface epithelium (OSE) not only has to undergo epithelial-mesenchymal transition (EMT), but also possess the properties of somatic stem-like cells. However, there is no evidence to indicate that both EMT alteration and stemness acquisition are linked to epithelial ovarian carcinomas.Methods In this study, we established a cell model of spontaneous oncogenic transformation of mouse OSE (MOSE). The cell proliferation was assessed using clonogenic survival and soft agar. The self-renewal of cancer stem-like cells (CSCs) was determined by spheroid culture. CD44 + /CD117 + cells were analyzed using flow cytometer. The PCR array was used to determine the EMT-related mRNA level. Expression of pan-keratin, vimentin, E-cadherin, Snail1 and Slug were detected using western-blotting and immunofluorescences, respectively. The tumorigenesis were monitored by limiting dilution assay in vitro and in vivo .Results Based on morphological change, chromosomal number and proliferating ability, we defined three sequential stages of transformed cells as early, intermediate and late MOSE cells, respectively. We found that MOSE cells had dual characteristics of not only epithelial but mesenchymal nature as well. Over time, MOSE cells spontaneously developed characteristics of malignant cells and generated tumor nodules expressing both Pan-keratin and Vimentin. Furthermore, we found that the neoplastic transformation of MOSE was accompanied by continuous EMT-inducing signals including Snail1 and Slug. Concurrently, the increase of CD44 + /CD117 + cells and their self-renewal ability were associated with the progression of spontaneous neoplastic transformation of MOSE cells in vitro. Conclusion These results indicated that both EMT alteration and stemness acquisition were closely correlated with the spontaneous malignant progression of MOSE cells. Our findings provide new insights into the future to combat epithelial ovarian carcinomas.

scholarly journals Spontaneous malignant transformation of ovarian surface epithelial cells correlates with EMT alteration and stemness acquisition

2019 ◽  
Author(s):  
Bingyan Li ◽  
Hemei Zhang ◽  
Yi Guo ◽  
Lizhi Liu ◽  
Yongfeng Hou ◽  
...  
Keyword(s):  
Epithelial Mesenchymal Transition ◽  
Cell Model ◽  
Mrna Level ◽  
Neoplastic Transformation ◽  
Surface Epithelium ◽  
Ovarian Carcinomas ◽  
Self Renewal ◽  
Mesenchymal Transition ◽  
Ovarian Surface

Abstract Background In order to complete the physiological function of postovulatory repair during repeated ovulation, the ovarian surface epithelium (OSE) not only has to undergo epithelial-mesenchymal transition (EMT), but also possess the properties of somatic stem-like cells. However, there is no evidence to indicate that both EMT alteration and stemness acquisition are linked to epithelial ovarian carcinomas.Methods In this study, we established a cell model of spontaneous oncogenic transformation of mouse OSE (MOSE). The cell proliferation was assessed using clonogenic survival and soft agar. The self-renewal of cancer stem-like cells (CSCs) was determined by spheroid culture. CD44 + /CD117 + cells were analyzed using flow cytometer. The PCR array was used to determine the EMT-related mRNA level. Expression of pan-keratin, vimentin, E-cadherin, Snail1 and Slug were detected using western-blotting and immunofluorescences, respectively. The tumorigenesis were monitored by limiting dilution assay in vitro and in vivo .Results Based on morphological change, chromosomal number and proliferating ability, we defined three sequential stages of transformed cells as early, intermediate and late MOSE cells, respectively. We found that MOSE cells had dual characteristics of not only epithelial but mesenchymal nature as well. Over time, MOSE cells spontaneously developed characteristics of malignant cells and generated tumor nodules expressing both Pan-keratin and Vimentin. Furthermore, we found that the neoplastic transformation of MOSE was accompanied by continuous EMT-inducing signals including Snail1 and Slug. Concurrently, the increase of CD44 + /CD117 + cells and their self-renewal ability were associated with the progression of spontaneous neoplastic transformation of MOSE cells in vitro. Conclusion These results indicated that both EMT alteration and stemness acquisition were closely correlated with the spontaneous malignant progression of MOSE cells. Our findings provide new insights into the future to combat epithelial ovarian carcinomas.

scholarly journals CircRNA PIP5K1A promotes the progression of glioma through upregulation of TCF12/PI3K/AKT pathway by sponging miR-515-5p

2020 ◽  
Author(s):  
Kebin Zheng ◽  
Haipeng Xie ◽  
Xiaosong Wu ◽  
Xichao Wen ◽  
Zhaomu Zeng ◽  
...  
Keyword(s):  
Epithelial Mesenchymal Transition ◽  
Cell Model ◽  
Luciferase Reporter ◽  
Akt Pathway ◽  
Circular Rnas ◽  
Rt Pcr ◽  
Mesenchymal Transition ◽  
Normal Tissues

Abstract BackgroundIncreasing studies have revealed that circular RNAs (CircRNAs) make great contribution to regulating tumor progression. Therefore, we intended to explore the expression characteristics, function, and related mechanisms of a novel type of circRNA, PIP5K1A in glioma. MethodsFirstly, RT-PCR was carried out to examine CircPIP5K1A expression in glioma tissues and adjacent normal tissues, and the correlation between CircPIP5K1A level and the clinical pathological indicators of glioma was analyzed. Then, the CircPIP5K1A expression in various glioma cell lines was detected, and a cell model of CircPIP5K1A overexpression and knockdown was constructed. Subsequently, cell proliferation and viability were detected by CCK8 method and BrdU staining, apoptosis was detected by flow cytometry, and cell invasion was examined by Transwell assay. The expression of TCF12, PI3K/AKT pathway apoptotic related proteins (including Caspase3, Bax and Bcl2) and epithelial-mesenchymal transition (EMT) markers (including E-cadherin, Vimentin and N-cadherin) by western blot or RT-PCR. ResultsThe results manifested that CircPIP5K1A was obviously upregulated in glioma tissues (compared with that in normal adjacent tissues), and overexpressed CircPIP5K1A was distinctly related to glioma volume and histopathological grade. Functionally, overexpressing CircPIP5K1A notably elevated the proliferation, invasion, EMT of glioma cells, and inhibited apoptosis both in vivo and in vitro. Besides, CircPIP5K1A also upregulated TCF12 and PI3K/AKT pathway activation. Bioinformatics analysis testified that miR-515-5p was a common target of CircPIP5K1A and TCF12, while dual luciferase reporter assay and RNA immunocoprecipitation (RIP) experiment further confirmed that CircPIP5K1A targeted miR-515-5p, which bound the 3'-untranslated region (UTR) of TCF12. ConclusionsAltogether, the study illustrated that CircPIP5K1A is a potential prognostic marker in glioma and regulates the development of glioma through the modulating miR-515-5p mediated TCF12/PI3K/AKT axis.

scholarly journals Downregulation of SLC27A6 by DNA Hypermethylation Promotes Proliferation but Suppresses Metastasis of Nasopharyngeal Carcinoma Through Modulating Lipid Metabolism

2022 ◽  
Vol 11 ◽  
Author(s):  
Xuemin Zhong ◽  
Yanping Yang ◽  
Bo Li ◽  
Pan Liang ◽  
Yiying Huang ◽  
...  
Keyword(s):  
Lipid Metabolism ◽  
Nasopharyngeal Carcinoma ◽  
Epithelial Mesenchymal Transition ◽  
Mrna Level ◽  
Promoter Hypermethylation ◽  
Migration And Invasion ◽  
Dna Hypermethylation ◽  
Mesenchymal Transition

Lipid is the building block and an important source of energy, contributing to the malignant behavior of tumor cells. Recent studies suggested that lipid droplets (LDs) accumulations were associated with nasopharyngeal carcinoma (NPC) progression. Solute carrier family 27 member 6 (SLC27A6) mediates the cellular uptake of long-chain fatty acid (LCFA), a necessary lipid component. However, the functions of SLC27A6 in NPC remain unknown. Here, we found a significant reduction of SLC27A6 mRNA in NPC tissues compared with normal nasopharyngeal epithelia (NNE). The promoter methylation ratio of SLC27A6 was greater in NPC than in non-cancerous tissues. The demethylation reagent 5-aza-2’-deoxycytidine (5-aza-dC) remarkably restored the mRNA expression of SLC27A6, suggesting that this gene was downregulated in NPC owing to DNA promoter hypermethylation. Furthermore, SLC27A6 overexpression level in NPC cell lines led to significant suppression of cell proliferation, clonogenicity in vitro, and tumorigenesis in vivo. Higher SLC27A6 expression, on the other hand, promoted NPC cell migration and invasion. In particular, re-expression of SLC27A6 faciliated epithelial-mesenchymal transition (EMT) signals in xenograft tumors. Furthermore, we observed that SLC27A6 enhanced the intracellular amount of triglyceride (TG) and total cholesterol (T-CHO) in NPC cells, contributing to lipid biosynthesis and increasing metastatic potential. Notably, the mRNA level of SLC27A6 was positively correlated with cancer stem cell (CSC) markers, CD24 and CD44. In summary, DNA promoter hypermethylation downregulated the expression of SLC27A6. Furthermore, re-expression of SLC27A6 inhibited the growth capacity of NPC cells but strengthened the CSC markers. Our findings revealed the dual role of SLC27A6 in NPC and shed novel light on the link between lipid metabolism and CSC maintenance.

scholarly journals CXCL14 Facilitates the Growth and Metastasis of Ovarian Carcinoma Cells via Activation of the Wnt/β-catenin Signaling Pathway

2021 ◽  
Author(s):  
Li-Na Gao ◽  
Man Hao ◽  
Xiao-Hui Liu ◽  
Li Zhang ◽  
Yan Dong ◽  
...  
Keyword(s):  
Cell Growth ◽  
Protein Expression ◽  
Signaling Pathway ◽  
Molecular Mechanisms ◽  
Epithelial Mesenchymal Transition ◽  
Mrna Level ◽  
Metastatic Potential ◽  
Mesenchymal Transition ◽  
Cell Invasiveness

Abstract Background There is an urgent need to identify potential targets in anticancer therapy to improve the survival and prognosis of patients with ovarian cancer (OC). Herein, we investigated the functional significance of chemokine (C-X-C motif) ligand 14 (CXCL14) in OC cell growth and epithelial–mesenchymal transition (EMT).MethodsqRT PCR and western blotting was used to detect CXCL14 mRNA level and protein expression, respectively. The functional mechanism of CXCL14 in OC was investigated by CCK-8, colony formation and transwell assays. The protein expression of CXCL14 and β-catenin in OC tissues was determined by immumohistochemical staining.ResultsWe demonstrated that high levels of CXCL14 were associated with a worse prognosis in patients with OC. CXCL14 knockdown considerably restrained the growth and invasion of OC cells in vitro. In contrast, ectopic CXCL14 overexpression yielded the opposite results. Investigations to determine the underlying molecular mechanisms revealed that the Wnt/β-catenin signaling pathway is involved in CXCL14-facilitated OC cell invasiveness.ConclusionThese data collectively demonstrate that CXCL14 contributes to OC cell growth and metastatic potential by regulating the Wnt/β-catenin signaling pathway.

scholarly journals Lgr5+CD44+EpCAM+ Strictly Defines Cancer Stem Cells in Human Colorectal Cancer

2018 ◽  
Vol 46 (2) ◽  
pp. 860-872 ◽  
Author(s):  
Zhengwei Leng ◽  
Qinghua Xia ◽  
Jinhuang Chen ◽  
Yong Li ◽  
Jiqian Xu ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Stem Cells ◽  
Cancer Stem Cells ◽  
Colony Formation ◽  
Epithelial Mesenchymal Transition ◽  
Tumor Xenograft ◽  
Self Renewal ◽  
Mesenchymal Transition

Background/Aims: Although EpCAM+CD44+ cells exhibit more stem-like properties than did EpCAM-CD44- cells, the specificity of EpCAM combined with CD44 in defining CSCs needs further improvement. Lgr5 is used as a biomarker to isolate cancer stem cells (CSCs) in colorectal cancer. However, it remains unclear whether Lgr5, along with EpCAM and CD44, can further identify and define CSCs in colorectal cancer. Methods: Lgr5+CD44+EpCAM+, Lgr5+CD44+EpCAM-, Lgr5+CD44-EpCAM+, Lgr5-CD44+EpCAM+, and Lgr5-CD44-EpCAM-cells were separately isolated using fluorescence-activated cell sorting (FACS). Colony formation, self-renewal, differentiation, and tumorigenic properties of these cells were investigated through in vitro experiments and in vivo tumor xenograft models. The expression of stemness genes and CSC- and epithelial-mesenchymal transition (EMT)-related genes, such as KLF4, Oct4, Sox2, Nanog, CD133, CD44, CD166, ALDH1, Lgr5, E-cadherin, ZO-1, Vimentin, Snail, Slug, and Twist, was examined using real-time PCR. Results: Lgr5-positive subpopulations exhibited higher capacities for colony formation, self-renewal, differentiation, and tumorigenicity as well as higher expression of stemness genes and mesenchymal genes and lower expression of epithelial genes than did Lgr5-negative subpopulations. Conclusion: Our data revealed that tumorigenic cells were highly restricted to Lgr5-positive subpopulations. Most importantly, Lgr5+CD44+EpCAM+ cells exhibited more pronounced CSC-like traits than did any other subpopulation, indicating that Lgr5 combined with CD44 and EpCAM can further improve the stem-like traits of CSCs in colorectal cancer.

Molecular pathways regulating EGF-induced epithelio-mesenchymal transition in human ovarian surface epithelium

2006 ◽  
Vol 290 (6) ◽  
pp. C1532-C1542 ◽  
Author(s):  
Nuzhat Ahmed ◽  
Sarah Maines-Bandiera ◽  
Michael A. Quinn ◽  
Waldemar G. Unger ◽  
Shoukat Dedhar ◽  
...  
Keyword(s):  
Molecular Mechanisms ◽  
Ovarian Surface Epithelium ◽  
Surface Epithelium ◽  
Ovarian Carcinomas ◽  
Physiological Basis ◽  
Mesenchymal Transition ◽  
Functional Changes ◽  
Akt Phosphorylation ◽  
Ovarian Surface ◽  
Gsk 3Β

The ovarian surface epithelium (OSE) is the precursor of common epithelial ovarian carcinomas. In the present study, we examined the molecular mechanisms and possible physiological basis for the propensity of OSE cells to undergo epithelio-mesenchymal transition (EMT) in response to environmental influences. We hypothesized that EMT may be a homeostatic mechanism that permits displaced OSE to assume a stromal phenotype within the ovarian cortex. We report that EGF in conjunction with hydrocortisone is the EMT-inducing factor of OSE as shown by changes to a fibroblast-like morphology and growth pattern. EGF increased cell motility, enhanced the activities of secreted pro-matrix metalloproteinase (MMP)-2 and -9, and enhanced expression and activation of Erk and integrin-linked kinase (ILK). Increased ILK expression correlated with the activation of PKB/Akt, the phosphorylation of GSK-3β, and the increased expression of cyclin E and cdk2 kinase. EGF withdrawal resulted in a more epithelial morphology and reversal of the EGF-induced activation of signaling pathways and pro-MMP activity. In contrast, treatment of EGF-treated cells with specific inhibitors of phosphatidylinositol 3-kinase, Mek, or ILK inhibited the inhibitor-specific pathways. The inhibitors caused suppression of EGF-induced migration and pro-MMP-2/-9 activities but did not lead to any change in EGF-induced mesenchymal morphology. ILK small interfering RNA inhibited Akt phosphorylation and reduced pro-MMP-2/-9 activities but had no effect on Erk activation or cell morphology. These results indicate that the EGF-induced morphological and functional changes in OSE cells are controlled by distinct signaling mechanisms working in concert. EMT of OSE cells displaced by ovulation likely permits their survival and integration with a fibroblast-like identity within the stroma. Failure to do so may lead to the formation of epithelium-derived inclusion cysts, which are known preferential sites of malignant transformation.

scholarly journals CXCL14 facilitates the growth and metastasis of ovarian carcinoma cells via activation of the Wnt/β-catenin signaling pathway

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Li-Na Gao ◽  
Man Hao ◽  
Xiao-Hui Liu ◽  
Li Zhang ◽  
Yan Dong ◽  
...  
Keyword(s):  
Cell Growth ◽  
Signaling Pathway ◽  
Molecular Mechanisms ◽  
Epithelial Mesenchymal Transition ◽  
Mrna Level ◽  
Metastatic Potential ◽  
Migration And Invasion ◽  
Migration Ability ◽  
Mesenchymal Transition

Abstract Background There is an urgent need to identify potential targets in anticancer therapy to improve the survival and prognosis of patients with ovarian cancer (OC). Herein, we investigated the functional significance of chemokine (C-X-C motif) ligand 14 (CXCL14) in OC cell growth and epithelial–mesenchymal transition (EMT). Methods qRT PCR and western blotting was used to detect CXCL14 mRNA level and protein expression, respectively. The functional mechanism of CXCL14 in OC was investigated by CCK-8, colony formation and transwell assays. The migration ability of OC cell was determined using wound healing. The protein expressions of CXCL14 and β-catenin in OC tissues were determined by immumohistochemical staining. Results We demonstrated that high levels of CXCL14 were associated with a worse prognosis in patients with OC. CXCL14 knockdown considerably restrained the growth, migration and invasion of OC cell in vitro. In contrast, ectopic CXCL14 overexpression yielded the opposite results. Investigations to determine the underlying molecular mechanisms revealed that the Wnt/β-catenin signaling pathway is involved in CXCL14-facilitated OC cell invasiveness. Conclusion These data collectively demonstrate that CXCL14 contributes to OC cell growth and metastatic potential by regulating the Wnt/β-catenin signaling pathway.

scholarly journals Phenotypic Plasticity of the Ovarian Surface Epithelium: TGF-β1 Induction of Epithelial to Mesenchymal Transition (EMT) in Vitro

Endocrinology ◽  
2010 ◽  
Vol 151 (11) ◽  
pp. 5497-5505 ◽  
Author(s):  
Yihong Zhu ◽  
Mikael Nilsson ◽  
Karin Sundfeldt
Keyword(s):  
Epithelial To Mesenchymal Transition ◽  
Ovarian Surface Epithelium ◽  
Surface Epithelium ◽  
Mesenchymal Transition ◽  
Epithelial Markers ◽  
Ovarian Surface ◽  
Epithelial Junction ◽  
Tgf Β1 ◽  
E Cadherin

Ovarian surface epithelium (OSE) is the most conceivable cell origin of epithelial ovarian carcinomas. Unlike many other epithelial tumors, the precancerous lesion acquires expression of epithelial markers, e.g. E-cadherin and claudins, suggesting that OSE cells undergo mesenchymal to epithelial transition (MET) during transformation. Recent findings indicate that TGF-β1, a prototypic stimulus of epithelial to mesenchymal transition (EMT), i.e. reverse to MET, is produced at significant amounts in the intact ovary. In the present study, we therefore investigated whether TGF-β1 changes the OSE phenotype accordingly, focusing on epithelial junction proteins and transcriptional EMT regulators quantified by real-time RT-PCR and Western blotting in cultured normal human OSE. Early OSE passages were found to paradoxically express de novo E-cadherin and also establish tight junctions exhibiting claudin-1 (but not claudin-3 and -4) and occludin. Stimulation with TGF-β1 (100 ng/ml) for 3–5 d down-regulated all these epithelial markers including Crumbs3 and also prevented the formation of an epithelial barrier This was accompanied by sustained expression of Snail and N-cadherin and transient expression of Slug, whereas Zeb1 (zinc finger E-box binding homeobox 1) and Twist mRNA levels were not significantly changed. In conclusion, TGF-β1 enforces the mesenchymal phenotype of OSE cells in vitro by an EMT-like process, leading to an altered molecular composition of the epithelial junction complex that partly coincides with the expression pattern of the native OSE. This suggests a potential role of TGF-β1-induced EMT in OSE under physiological conditions and possibly also in epithelial ovarian tumorigenesis.

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