Preliminary identification of endometrial cancer stem cells in vitro and in vivo

Kong, Fanfei; Li, Da; Yang, Hui; Ma, Jian; Pan, Xin; Liu, Hongxiang; Huo, Jianing; Ma, Xiaoxin

doi:10.1016/j.bbrc.2017.06.070

Cited by 17 publications

(21 citation statements)

References 31 publications

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“…Under normoxic conditions, cells were cultured in an incubator with 5% CO 2 and 95% air (21% O 2 ). Under hypoxic conditions, cells were seeded in a modular incubator chamber (Billups-Rothenberg) and flushed for 2 min at 2 psi with a gas mixture containing 2% O 2 , 5% CO 2 and 94% N 2 63 . During culture, ECSCs were treated with CoCl 2 (7646-79-9, Sigma-Aldrich, St. Louis, MO, USA) or KC7F2 (HY-18777, MedChemExpress, Monmouth Junction, NJ, USA) as noted in the experiments.…”

Section: Methodsmentioning

confidence: 99%

Hypoxia induces an endometrial cancer stem-like cell phenotype via HIF-dependent demethylation of SOX2 mRNA

et al. 2020

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Endometrial cancer stem cells (ECSCs) are stem-like cells endowed with self-renewal and differentiation abilities, and these cells are essential for cancer progression in endometrial cancer (EC). As hallmarks of the tumour microenvironment (TME), hypoxia and hypoxia-inducing factors (HIFs) give rise to the dysregulation of tumour stemness genes, such as SOX2. Against this backdrop, we investigated the regulatory mechanisms regulated by HIFs and SOX2 in ECSCs during EC development. Here, ECSCs isolated from EC cell lines and tissues were found to express stemness genes (CD133 and aldehyde dehydrogenase, ALDH1) following the induction of their ECSC expansion. Notably, m6A methylation of RNA and HIF-1α/2α-dependent AlkB homologue 5 (ALKBH5) participate in the regulation of HIFs and SOX2 in EC, as confirmed by the observations that mRNA levels of m6A demethylases and ALKBH5 significantly increase under hypoxic conditions in ECSCs. Moreover, hypoxia and high ALKBH5 levels restore the stem-like state of differentiated ECSCs and increase the ECSC-like phenotype, whereas the knockdown of HIFs or ALKBH5 significantly reduces their tumour initiation capacity. In addition, our findings validate the role of ALKBH5 in promoting SOX2 transcription via mRNA demethylation, thereby maintaining the stem-like state and tumorigenicity potential of ECSCs. In conclusion, these observations demonstrate a critical role for m6A methylation-mediated regulation of the HIF-ALKBH5-SOX2 axis during ECSC expansion in hypoxic TMEs.

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Section: Methodsmentioning

confidence: 99%

Hypoxia induces an endometrial cancer stem-like cell phenotype via HIF-dependent demethylation of SOX2 mRNA

et al. 2020

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“…Indeed, different approaches have been developed, among which fluorescence activated cell sorting, individuation of SP, with and without the use of Hoechst 33342, and use of a serum free suspension cultivation with the addition of growth factors. Specifically, this medium drives to formation of microsphere, proliferation of CSCs and elimination of differentiated cells [86].…”

Section: Cscs In Endometrial Cancer: Role and Biomarkersmentioning

confidence: 99%

Endometrial Cancer Stem Cells: Role, Characterization and Therapeutic Implications

Giannone

Attademo

Scotto

et al. 2019

Cancers

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Endometrial cancer (EC) is the most frequent gynecological cancer. In patients with relapsed and advanced disease, prognosis is still dismal and development of resistance is common. In this context, endometrial Cancer Stem Cells (eCSC), stem-like cells capable to self-renewal and differentiation in mature cancer cells, represent a potential field of expansion for drug development. The aim of this review is to characterize the role of eCSC in EC, their features and how they could be targeted. CSC are involved in progression, invasiveness and metastasis (though epithelial to mesenchimal transition, EMT), as well as chemoresistance in EC. Nevertheless, isolation of eCSC is still controversial. Indeed, CD133, Aldheyde dehydrogenase (ALDH), CD117, CD55 and CD44 are enriched in CSCs but there is no universal marker nowadays. The most frequently activated pathways in eCSC are Wingless-INT (Wnt)/β-catenin, Notch1, and Hedghog, with a high expression of self-renewal transcription factors like Octamer binding transcription factor 4 (OCT), B Lymphoma Mo-MLV Insertion Region 1 Homolog (BMI1), North American Network Operations Group Homebox protein (NANOG), and SRY-Box 2 (SOX2). These pathways have been targeted with selective drugs alone or in combination with chemotherapy and immunotherapy. Unfortunately, although preclinical results are encouraging, few clinical data are available.

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“…According to our previous research results [7] , we performed ow cytometry sorting assay. After Ishikawa cells were cultured and passaged in serum-free medium, the cell pellet was collected and washed once with phosphate-buffered saline (PBS).…”

Section: Flow Cytometry Sorting Assaymentioning

confidence: 99%

“…CSCs can initiate tumor formation and promote tumor cell proliferation, while differentiation of component tumor cells plays a vital role in the occurrence, development, metastasis, recurrence, and drug resistance of malignant tumors [5,6] . In our previous research, we used serumfree suspension culture to isolate endometrial cancer stem cells (ECSCs) from Ishikawa cells [7] .…”

Section: Introductionmentioning

confidence: 99%

LncRNA MONC suppresses the malignant phenotype of Endometrial Cancer Stem Cells and Endometrial Carcinoma Cells by regulating the MiR-636/GLCE axis

Huo

et al. 2021

Preprint

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Background Emerging evidence shows that abnormal expression of long non-coding RNA is involved in the occurrence and development of various tumors. LncRNA MONC is abnormally expressed in head and neck squamous cell carcinoma, lung cancer, colorectal cancer, and acute megakaryocytic leukemia, but the biological function and potential regulatory mechanism of MONC in endometrial cancer stem cells (ECSCs) and endometrial cancer cells (ECCs) have not been studied. Methods We used qRT-PCR to detect the expression of MONC, miR-636 and GLCE in normal human endometrial tissues and endometrial carcinoma tissues. Luciferase assay was used to verify the binding sites between MONC and miR-636 and between miR-636 and GLCE. The double fluorescence in situ hybridization was used to locate MONC and miR-636 in cells. Endometrial cancer stem cells were obtained by Flow cytometry sorting assay. Sphere formation assay, CCK-8 assay, transwell invasion assay, cell cycle analysis and apoptosis assay were used to detect the effects of MONC/miR-636/GLCE axis on the malignant biological behavior of ECSCs and endometrial carcinoma cells (ECCs). The effect of MONC on the epithelial-to-mesenchymal transition (EMT) process was detected using western blot. Finally, we conducted in vivo verification through Tumor xenografts in nude mice. Results In this study, we aimed to explore the tumor suppressive effect and mechanism of MONC in regulating ECSCs and ECCs. We found MONC is low expression in endometrial carcinoma (EC), MONC and miR-636 are relatively co-localized in the cytoplasm. MONC directly inhibits the malignant biological behavior of ECSCs and ECCs by directly inhibiting miR-636. Simultaneously, miR-636 may indirectly reduce the expression of MONC. Down-regulation of miR-636 may promote GLCE expression by targeting the 3'-untranslated region (UTR) of the downstream gene GLCE, thereby inhibiting the progression of ECSCs. MONC combined with miR-636 inhibited tumor epithelial-to-mesenchymal transition (EMT) process. In addition, we verified the tumor suppressive effect of MONC in nude mice, miR-636 can rescue the tumor suppressive effect of overexpressing MONC. Conclusions In conclusion, this study shows that MONC inhibits the malignant phenotypes of ECSCs and ECCs by regulating the miR-636/GLCE axis. The MONC/miR-636/GLCE axis may provide novel treatment avenues for human EC.

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Preliminary identification of endometrial cancer stem cells in vitro and in vivo

Cited by 17 publications

References 31 publications

Hypoxia induces an endometrial cancer stem-like cell phenotype via HIF-dependent demethylation of SOX2 mRNA

Hypoxia induces an endometrial cancer stem-like cell phenotype via HIF-dependent demethylation of SOX2 mRNA

Endometrial Cancer Stem Cells: Role, Characterization and Therapeutic Implications

LncRNA MONC suppresses the malignant phenotype of Endometrial Cancer Stem Cells and Endometrial Carcinoma Cells by regulating the MiR-636/GLCE axis

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