Celastrus orbiculatus Extract Reduces Stemness of Gastric Cancer Stem Cells by Targeting PDCD4 and EIF3H

Background Celastrus orbiculatus ethyl acetate extract (COE) has shown a strong anti-gastric cancer effect, but the understanding of its mechanism is still lacking. The results of previous studies indicated that COE may be able to inhibit the stemness of gastric cancer stem cells (GCSCs) by regulating PDCD4 and EIF3H expression. Aims To explore if COE could inhibit the stemness of GCSCs by regulating PDCD4 and EIF3H expression in vitro and in vivo. Procedure The GCSCs model was established by stem cell-conditioned culture. Spheroid formation and flow cytometry assays were used to detect the effect of COE on the spheroid formation ability of GCSCs and the percentage of CD44+/CD24+ and ALDH+ cell subpopulations. Western blot analysis was applied to measure the expression of GCSCs biomarkers (Nanog, Oct-4, and SOX-2), PDCD4, and EIF3H in GCSCs treated with COE; and RT-PCR was performed to investigate the effect of COE on PDCD4 mRNA expression in GCSCs. An in vivo tumorigenicity experiment was also conducted to evaluate the effect of COE on tumor-initiating ability of GCSCs in vivo; and the expression of PDCD4 and EIF3H in xenograft tissues was examined by immunohistochemistry (IHC) staining. Results After culture in stem cell-conditioned medium, SGC7901 cells manifested significantly enhanced spheroid formation ability, upregulated Nanog, Oct-4, and SOX-2 expression and increased percentages of CD44+/CD24+ and ALDH+ cell subpopulations, indicating successful establishment of the GCSCs model. COE treatment significantly inhibited the spheroid formation ability of GCSCs and reduced the percentage of CD44+/CD24+ and ALDH+ cell subpopulations. The western blot analysis showed a significant decrease of Nanog, Oct-4, SOX-2, and EIF3H expression and an increase of PDCD4 expression in GCSCs after COE treatment in a concentration-dependent manner. COE treatment also significantly upregulated the mRNA expression of PDCD4 in GCSCs. In addition, COE displayed a strong inhibitory effect on the tumor-initiating ability of GCSCs in vivo and upregulated PDCD4 and downregulated EIF3H expression in xenograft tissues. Conclusion COE may be able to inhibit GC growth by suppressing the stemness of GCSCs via regulating PDCD4 and EIF3H expression.

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Gastric Cancer Stem Cells Survive in Stress Environments via Their Autophagy System

10.21203/rs.3.rs-91127/v1 ◽

2020 ◽

Author(s):

Shingo Togano ◽

Masakazu Yashiro ◽

Go Masuda ◽

Atsushi Sugimoto ◽

Yuichiro Miki ◽

...

Keyword(s):

Gastric Cancer ◽

Stem Cells ◽

Stem Cell ◽

Electron Microscope ◽

Cancer Stem Cells ◽

Western Blot ◽

Stem Cell Markers ◽

Cell Markers ◽

Autophagy Inhibitor ◽

Stress Environments

Abstract Background: It has been reported that cancer stem cells (CSCs) play an important role in the progression of carcinoma and have a high potential for survival in various stress environments such as starvation and hypoxia. However, the mechanisms responsible for the capacity of CSCs to survive under stresses have been unclear. The aim of this study was to clarify the significance of the autophagy systems of CSCs under stress environments.Methods: Four human gastric cancer cell line, OCUM-12, OCUM-2MD3, MKN-45 and MKN-74 were used. Side population (SP) cells were sorted from the parent OCUM-12 and OCUM-2MD3, as CSC rich cells. The expression of stem cell markers was examined by RT-PCR. The viability of cancer cells under starvation and hypoxia (1%O2) was evaluated by MTT assay with or without the autophagy inhibitor, chloroquine. The expression level of the autophagy molecule LC3-II was examined by western blot. The numbers of autophagosomes and autolysosomes were counted by electron microscope.Results: SP cells of OCUM-12 showed a higher expression of stem cell markers and higher viability in starvation and hypoxia. Western blot and electron microscope examinations indicated that the autophagy was more induced in SP cells than in parent cells. The autophagy inhibitor significantly decreased the viability and the proportion of SP cells under the stress environments.Conclusions: Cancer stem cells of gastric cancer might maintain their viability under stress environments of starvation and hypoxia via the autophagy system. Autophagy inhibitors might be a promising therapeutic agent for gastric cancer.

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MicroRNA-34 dysregulation in gastric cancer and gastric cancer stem cell

Tumor Biology ◽

10.1177/1010428317701652 ◽

2017 ◽

Vol 39 (5) ◽

pp. 101042831770165 ◽

Cited By ~ 14

Author(s):

Narjes Jafari ◽

Saeid Abediankenari

Keyword(s):

Gastric Cancer ◽

Stem Cells ◽

Stem Cell ◽

Cancer Stem Cells ◽

Cancer Stem Cell ◽

Patient Treatment ◽

Small Subset ◽

Aberrant Expression ◽

Wide Range ◽

Gastric Cancer Stem Cells

Gastric cancer is a major cause of cancer mortality worldwide, with a low survival rate for patients with advanced forms of the disease. Over the recent decades, the investigation of the pathophysiological mechanisms of tumourigenesis has opened promising avenues to understand some of the complexities of cancer treatment. However, tumour regeneration and metastasis impose great difficulty for gastric cancer cure. In recent years, cancer stem cells – a small subset of tumour cells in many cancers – have become a major focus of cancer research. Cancer stem cells are capable of self-renewal and are known to be responsible for tumour initiation, metastasis, therapy resistance and cancer recurrence. Recent studies have revealed the key role of microRNAs – small noncoding RNAs regulating gene expression – in these processes. MicroRNAs play crucial roles in the regulation of a wide range of biological processes in a post-transcriptional manner, though their expression is dysregulated in most malignancies, including gastric cancer. In this article, we review the consequences of aberrant expression of microRNA-34 in cancer and cancer stem cells, with a specific focus on the miR-34 dysregulation in gastric cancer and gastric cancer stem cells. We address the critical effects of the aberrant expression of miR-34 and its target genes in maintaining cancer stem cell properties. Information collection and discussion about the advancements in gastric cancer stem cells and microRNAs can be useful for providing novel insights into patient treatment.

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Gastric cancer stem cells survive in stress environments via their autophagy system

Scientific Reports ◽

10.1038/s41598-021-00155-3 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Shingo Togano ◽

Masakazu Yashiro ◽

Go Masuda ◽

Atsushi Sugimoto ◽

Yuichiro Miki ◽

...

Keyword(s):

Gastric Cancer ◽

Stem Cells ◽

Stem Cell ◽

Electron Microscope ◽

Cancer Stem Cells ◽

Western Blot ◽

Gastric Cancer Cell Line ◽

Stem Cell Markers ◽

Cell Markers ◽

Stress Environments

AbstractCancer stem cells (CSCs) play an important role in the progression of carcinoma and have a high potential for survival in stress environments. However, the mechanisms of survival potential of CSCs have been unclear. The aim of this study was to clarify the significance of autophagy systems of CSCs under stress environments. Four gastric cancer cell line were used. Side population (SP) cells were sorted from the parent cells, as CSC rich cells. The expression of stem cell markers was examined by RT-PCR. The viability of cancer cells under starvation and hypoxia was evaluated. The expression level of the autophagy molecule LC3B-II was examined by western blot. The numbers of autophagosomes and autolysosomes were counted by electron microscope. SP cells of OCUM-12 showed a higher expression of stem cell markers and higher viability in starvation and hypoxia. Western blot and electron microscope examinations indicated that the autophagy was more induced in SP cells than in parent cells. The autophagy inhibitor significantly decreased the viability under the stress environments. These findings suggested that Cancer stem cells of gastric cancer might maintain their viability via the autophagy system. Autophagy inhibitors might be a promising therapeutic agent for gastric cancer.

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In vitro and in vivo effects of miRNA-19b/20a/92a on gastric cancer stem cells and the related mechanism

International Journal of Medical Sciences ◽

10.7150/ijms.21164 ◽

2018 ◽

Vol 15 (1) ◽

pp. 86-94 ◽

Cited By ~ 8

Author(s):

Qianwen Shao ◽

Jing Xu ◽

Xin Guan ◽

Bing Zhou ◽

Wei Wei ◽

...

Keyword(s):

Gastric Cancer ◽

Stem Cells ◽

Cancer Stem Cells ◽

Gastric Cancer Stem Cells ◽

In Vivo Effects

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Requirement of transcription factor NME2 for the maintenance of the stemness of gastric cancer stem-like cells

Cell Death and Disease ◽

10.1038/s41419-021-04234-1 ◽

2021 ◽

Vol 12 (10) ◽

Author(s):

Yaxin Qi ◽

Jun Wei ◽

Xiaobo Zhang

Keyword(s):

Gastric Cancer ◽

Stem Cells ◽

Transcription Factor ◽

Transcription Factors ◽

Cancer Stem Cells ◽

Nucleoside Diphosphate Kinase ◽

Gastric Cancer Stem Cells ◽

New Perspective

AbstractCancer stem cells (CSCs), which can self-renew and produce heterogeneous cancer cells, are the key factors during tumorigenesis. Transcription factors take essential effects on CSCs. However, the role of transcription factors in regulating the stemness of gastric cancer stem-like cells has not been well explored. In this investigation, it was found that transcription factor NME2 (NME/NM23 nucleoside diphosphate kinase 2) was upregulated in gastric cancer stem-like cells that sorted from the solid tumors of patients with gastric cancer and gastric cancer cell lines. NME2 could preserve the stemness of gastric cancer stem-like cells via suppressing their apoptosis. In vitro and in vivo data revealed that NME2 was crucial for maintaining the stemness of gastric cancer stem cells by enhancing the expression of anti-apoptosis genes. Consequently, our data contributed a new perspective to the relationship between transcription factor and the stemness maintenance of gastric cancer stem cells.

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Chemotoxicity-induced exosomal lncFERO regulates ferroptosis and stemness in gastric cancer stem cells

Cell Death and Disease ◽

10.1038/s41419-021-04406-z ◽

2021 ◽

Vol 12 (12) ◽

Author(s):

Haiyang Zhang ◽

Meng Wang ◽

Yi He ◽

Ting Deng ◽

Rui Liu ◽

...

Keyword(s):

Gastric Cancer ◽

Stem Cells ◽

Cell Death ◽

Cancer Stem Cells ◽

In Vivo Experiments ◽

Gastric Cancer Stem Cells ◽

Nuclear Ribonucleoprotein ◽

Sphere Formation

AbstractCancer stem cells (CSCs) are an important cause of tumor recurrence and drug resistance. As a new type of cell death that relies on iron ions and is strictly regulated by intracellular and extracellular signals, the role of ferroptosis in tumor stem cells deserves extensive attention. Mass spectrum was applied to screen for ferroptosis-related proteins in gastric cancer (GC). Sphere-formation assay was used to estimate the stemness of gastric cancer stem cells (GCSCs). Exosomal lnc-ENDOG-1:1 (lncFERO) was isolated by ultracentrifugation. Ferroptosis was induced by erastin and was assessed by detecting lipid ROS, mitochondrial membrane potential, and cell death. Furthermore, a series of functional in vitro and in vivo experiments were conducted to evaluate the effects of lncFERO on regulating ferroptosis and chemosensitivity in GCSCs. Here, we showed that stearoyl-CoA-desaturase (SCD1) played a key role in regulating lipid metabolism and ferroptosis in GCSCs. Importantly, exosomal lncFERO (exo-lncFERO) derived from GC cells was demonstrated to promote SCD1 expression by directly interacting with SCD1 mRNA and recruiting heterogeneous nuclear ribonucleoprotein A1 (hnRNPA1), which resulted in the dysregulation of PUFA levels and the suppression of ferroptosis in GCSCs. Moreover, we found that hnRNPA1 was also involved in lncFERO packing into exosomes in GC cells, and both in vitro and in vivo data suggested that chemotoxicity induced lncFERO secretion from GC cells by upregulating hnRNPA1 expression, leading to enhanced stemness and acquired chemo-resistance. All these data suggest that GC cells derived exo-lncFERO controls GCSC tumorigenic properties through suppressing ferroptosis, and targeting exo-lncFERO/hnRNPA1/SCD1 axis combined with chemotherapy could be a promising CSC-based strategy for the treatment of GC.

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Increased Production of Matrix Metalloproteinases in Helicobacter pylori Infection that Stimulates Gastric Cancer Stem Cells

International Journal of Veterinary Science ◽

10.37422/ijvs/20.043 ◽

2020 ◽

Keyword(s):

Gastric Cancer ◽

Stem Cells ◽

Helicobacter Pylori ◽

Cancer Stem Cells ◽

Histopathological Examination ◽

Physiological Saline ◽

Rrna Gene ◽

Peptic Ulcers ◽

Gastric Cancer Stem Cells ◽

H Pylori

Background and aim: Helicobacter pylori (H. pylori) is an incriminated pathogen causing diseases in both animals and humans and considered a zoonotic pathogen. H. pylori infection is considered a cause of gastric cancer, which rests a significant health care challenge. This study analyzes the expression pattern of matrix metalloprotein 2 (MMP-2) in patients with Helicobacter pylori-associated gastritis and the effect of H. pylori on gastric cancer stem cells, as well as study the role of helicon bacteriosis in dog in transmission of H. pylori infection to human. Materials and methods: Fifty-five of each sample (gastric biopsy, blood and stool) were collected from patients suffering from dyspepsia, chronic vomiting and perforated peptic ulcers and also from apparent healthy dogs. The investigation detected H. pylori by serological and histopathological examination. Biopsies were stored in physiological saline for identification of H. pylori by conventional time PCR. MMP-2 and Gastric cancer stem cells were then identified by immunohistochemistry. Results: Serological identification for H. pylori Antigen and Antibodies revealed (63% human, 50% dogs) and (87% human, 90% dogs) respectively were positive. Genotyping of H. pylori based on 16S rRNA gene showed 54.5% of human and 35% of dogs were positive. Immunohistochemistry revealed strong expression of CD44 in H. pylori- associated gastric cancer cases, MMP-2 expression was observed in all neoplastic lesions associated with H. pylori infection. Conclusion: H. pylori infection affects gastric mucosa and induces changes in gastric stem cells altering their differentiation and increased expression of MMP’s and CD44with a resultant potentiation of oncogenic alteration. In addition the up-regulation of both markers could be an instrumental to interpret the origination of gastric cancer.

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