MiR-29b may suppresses peritoneal metastases through inhibition of the mesothelial–mesenchymal transition (MMT) of human peritoneal mesothelial cells

AbstractPeritoneal dissemination is a major metastatic pathway for gastrointestinal and ovarian malignancies. The miR-29b family is downregulated in peritoneal fluids in patients with peritoneal metastases (PM). We examined the effect of miR-29b on mesothelial cells (MC) which play critical a role in the development of PM through mesothelial-mesenchymal transition (MMT). Human peritoneal mesothelial cells (HPMCs) were isolated from surgically resected omental tissue and MMT induced by stimulation with 10 ng/ml TGF-β1. MiR-29b mimics and negative control miR were transfected by lipofection using RNAiMAX and the effects on the MMT evaluated in vitro. HPMC produced substantial amounts of miR-29b which was markedly inhibited by TGF-β1. TGF-β1 stimulation of HPMC induced morphological changes with decreased expression of E-cadherin and calretinin, and increased expression of vimentin and fibronectin. TGF-β1 also enhanced proliferation and migration of HPMC as well as adhesion of tumor cells in a fibronectin dependent manner. However, all events were strongly abrogated by simultaneous transfection of miR-29b. MiR-29b inhibits TGF-β1 induced MMT and replacement of miR-29b in the peritoneal cavity might be effective to prevent development of PM partly through the effects on MC.

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MiR-29b suppresses peritoneal metastases through inhibition of the mesothelial-mesenchymal transition (MMT) of human peritoneal mesothelial cells

10.21203/rs.3.rs-774286/v1 ◽

2021 ◽

Author(s):

Yuki Kimura ◽

Hideyuki Ohzawa ◽

Hideyo Miyato ◽

Yuki Kaneko ◽

Kazuya Takahashi ◽

...

Keyword(s):

Morphological Changes ◽

Negative Control ◽

Mesothelial Cells ◽

Peritoneal Metastases ◽

Dependent Manner ◽

Mesenchymal Transition ◽

Peritoneal Mesothelial Cells ◽

Human Peritoneal Mesothelial Cells ◽

And Migration

Abstract Background: Peritoneal dissemination is a major metastatic pathway for gastrointestinal and ovarian malignancies. The miR-29b family is downregulated in peritoneal fluids in patients with peritoneal metastases (PM). We examined the effect of miR-29b on mesothelial cells (MC) which play critical a role in the development of PM through mesothelial-mesenchymal transition (MMT). Methods: Human peritoneal mesothelial cells (HPMCs) were isolated from surgically resected omental tissue and MMT induced by stimulation with 10 ng/ml TGF-b1. MiR-29b mimics and negative control miR were transfected by lipofection using RNAiMAX and the effects on the MMT evaluated in vitro. Results: HPMC produced substantial amounts of miR-29b which was markedly inhibited by TGF-b1. TGF-b1 stimulation of HPMC induced morphological changes with decreased expression of E-cadherin and calretinin, and increased expression of vimentin and fibronectin. TGF-b1 also enhanced proliferation and migration of HPMC as well as adhesion of tumor cells in a fibronectin dependent manner. However, all events were strongly abrogated by simultaneous transfection of miR-29b. Conclusion: MiR-29b inhibits TGF-b1 induced MMT and replacement of miR-29b in the peritoneal cavity might be effective to prevent development of PM partly through the effects on MC.

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miRNA589 Regulates Epithelial-Mesenchymal Transition in Human Peritoneal Mesothelial Cells

Journal of Biomedicine and Biotechnology ◽

10.1155/2012/673096 ◽

2012 ◽

Vol 2012 ◽

pp. 1-6 ◽

Cited By ~ 15

Author(s):

Ke Zhang ◽

Hao Zhang ◽

Xun Zhou ◽

Wen-bin Tang ◽

Li Xiao ◽

...

Keyword(s):

Epithelial Mesenchymal Transition ◽

Mesothelial Cell ◽

Mesothelial Cells ◽

Control Group ◽

Peritoneal Fibrosis ◽

Mesenchymal Transition ◽

Peritoneal Mesothelial Cells ◽

Human Peritoneal Mesothelial Cells ◽

E Cadherin

Background. microRNA (miRNA, miR) are thought to interact with multiple mRNAs which are involved in the EMT process. But the role of miRNAs in peritoneal fibrosis has remained unknown.Objective. To determine if miRNA589 regulates the EMT induced by TGFβ1 in human peritoneal mesothelial cell line (HMrSV5 cells).Methods. 1. Level of miR589 was detected in both human peritoneal mesothelial cells (HPMCs) isolated from continuous ambulatory peritoneal dialysis (CAPD) patients’ effluent and HMrSV5 cells treated with or without TGFβ1. 2. HMrSV5 cells were divided into three groups: control group, TGFβ1 group, and pre-miR-589+TGFβ1 group. The level of miRNA589 was determined by realtime PCR. The expressions of ZO-1, vimentin, and E-cadherin in HPMCs were detected, respectively.Results. Decreased level of miRNA589 was obtained in either HPMCs of long-term CAPD patients or HMrSV5 cells treated with TGFβ1. In vitro, TGFβ1 led to upregulation of vimentin and downregulation of ZO-1 as well as E-cadherin in HMrSV5 cells, which suggested EMT, was induced. The changes were accompanied with notably decreased level of miRNA589 in HMrSV5 cells treated with TGFβ1. Overexpression of miRNA589 by transfection with pre-miRNA589 partially reversed these EMT changes.Conclusion. miRNA589 mediates TGFβ1 induced EMT in human peritoneal mesothelial cells.

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Interleukin-1β Stimulates Glucose Uptake of Human Peritoneal Mesothelial Cells In Vitro

Peritoneal Dialysis International ◽

10.1177/089686089601601s09 ◽

1996 ◽

Vol 16 (1_suppl) ◽

pp. 58-60 ◽

Cited By ~ 5

Author(s):

Michael Kruse ◽

Arezki Mahiout ◽

Volker Kliem ◽

Peter Kurz ◽

Karl-Martin Koch ◽

...

Keyword(s):

Glucose Uptake ◽

Glucose Transporter ◽

Cytochalasin B ◽

Interleukin 1 ◽

Glucose Transporters ◽

Mesothelial Cells ◽

Dependent Manner ◽

Peritoneal Mesothelial Cells ◽

Human Peritoneal Mesothelial Cells

To investigate whether the glucose uptake (GU) of human peritoneal mesothelial cells (HPMC) is mediated by glucose transporters and whether this uptake is influenced by interleukin 1–β (IL-1β), we measured 2-deoxy-(3H)-GU of HPMC in vitro, after exposing the cells for different times (two and 12 hours) to increasing concentrations (0.1, 1.0, and 2.0 ng/mL) of IL-1 β. To exclude a noncarrier-mediated transport, GU was also tested in the presence of cytochalasin B. All experiments were performed in triplicate in the cells of two donors. Cytochalasin B inhibits GU of HPMC almost completely. GU of HPMC is not stimulated by insulin. GU is stimulated by IL-1 β in a dose-dependent manner. These data indicate a GU of HPMC, which is mediated by a glucose transporter and stimulated by IL-1 β. The increased uptake of glucose from the dialysate In patients with peritonitis may be mediated by a (cytokineinduced) increased activity of HPMC glucose transporters.

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Glycerol Toxicity for Human Peritoneal Mesothelial Cells in Culture: Comparison with Glucose

The International Journal of Artificial Organs ◽

10.1177/039139889401700502 ◽

1994 ◽

Vol 17 (5) ◽

pp. 252-260 ◽

Cited By ~ 14

Author(s):

J. Witowski ◽

J. Knapowski

Keyword(s):

Mesothelial Cell ◽

Cellular Protein ◽

Mesothelial Cells ◽

Dependent Manner ◽

Peritoneal Mesothelial Cells ◽

Proline Incorporation ◽

Human Peritoneal Mesothelial Cells ◽

Dose Dependent Decrease ◽

Dose Dependent

Glycerol has been proposed as a substitute osmotic agent for glucose in peritoneal dialysis fluids. We have compared the effect of glycerol and glucose on the function of human peritoneal mesothelial cells (HPMC) in vitro. The viability of HPMC was not affected by glycerol (up to 250 mM), whereas it was reduced by glucose in a time- and dose-dependent manner, as assessed by the LDH release. Although the incubation of HPMC with glycerol induced a dose-dependent decrease in HPMC proliferation, the effect was significantly less inhibitory than that produced by glucose. In HPMC treated with 90 mM of glycerol or glucose the incorporation of [3H]-thymidine had reached 79.0±19.3% and 55.3+4.0% of the control (p<0.05 and p<0.01), respectively. As measured by the [methyl-14C]-choline incorporation, the intracellular amount of newly synthesized phospholipids was reduced from (cpm/μg cellular protein) 147±58 in control HPMC to 59+15 in cells exposed to 90 mM of glucose (p<0.01), but not affected by glycerol (163±65). On the other hand, both glycerol and glucose (90 mM) decreased the synthesis of proteins (as assessed by the [3H]-proline incorporation) and interfered with potassium (86Rb) transport mechanisms in HPMC. Our data suggest that there exist some possibly advantageous aspects of glycerol as far as mesothelial cell biocompatibility profile is concerned.

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Correlation between the donor age and the proliferative lifespan of human peritoneal mesothelial cells in vitro: Is TGF-β1 a link?

Experimental Gerontology ◽

10.1016/j.exger.2007.04.012 ◽

2007 ◽

Vol 42 (8) ◽

pp. 840-843 ◽

Cited By ~ 10

Author(s):

Krzysztof Książek ◽

Marek Winckiewicz ◽

Ryszard Staniszewski ◽

Andrzej Bręborowicz ◽

Janusz Witowski

Keyword(s):

Mesothelial Cells ◽

Donor Age ◽

Peritoneal Mesothelial Cells ◽

Human Peritoneal Mesothelial Cells ◽

Proliferative Lifespan ◽

Tgf Β1

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Activation of General Control Nonderepressible-2 Kinase Ameliorates Glucotoxicity in Human Peritoneal Mesothelial Cells, Preserves Their Integrity, and Prevents Mesothelial to Mesenchymal Transition

Biomolecules ◽

10.3390/biom9120832 ◽

2019 ◽

Vol 9 (12) ◽

pp. 832

Author(s):

Theodoros Eleftheriadis ◽

Georgios Pissas ◽

Georgia Antoniadi ◽

Evdokia Nikolaou ◽

Spyridon Golfinopoulos ◽

...

Keyword(s):

High Glucose ◽

Cell Apoptosis ◽

Glucose Transporter ◽

Mesothelial Cells ◽

General Control ◽

Mesenchymal Transition ◽

Peritoneal Mesothelial Cells ◽

Ultrafiltration Failure ◽

Human Peritoneal Mesothelial Cells ◽

Tgf Β1

Along with infections, ultrafiltration failure due to the toxicity of glucose-containing peritoneal dialysis (PD) solutions is the Achilles’ heel of PD method. Triggered by the protective effect of general control nonderepressible-2 (GCN-2) kinase activation against high-glucose conditions in other cell types, we evaluated whether the same occurs in human peritoneal mesothelial cells. We activated GCN-2 kinase with halofuginone or tryptophanol, and assessed the impact of this intervention on glucose transporter-1, glucose transporter-3, and sodium-glucose cotransporter-1, glucose influx, reactive oxygen species (ROS), and the events that result in glucotoxicity. These involve the inhibition of glyceraldehyde 3-phosphate dehydrogenase and the diversion of upstream glycolytic products to the aldose pathway (assessed by D-sorbitol), the lipid synthesis pathway (assessed by protein kinase C activity), the hexosamine pathway (determined by O-linked β-N-acetyl glucosamine-modified proteins), and the advanced glycation end products generation pathway (assessed by methylglyoxal). Then, we examined the production of the profibrotic transforming growth factor-β1 (TGF-β1), the pro-inflammatory interleukin-8 (IL-8). Cell apoptosis was assessed by cleaved caspase-3, and mesothelial to mesenchymal transition (MMT) was evaluated by α-smooth muscle actin protein. High-glucose conditions increased glucose transporters, glucose influx, ROS, all the high-glucose-induced harmful pathways, TGF-β1 and IL-8, cell apoptosis, and MMT. Halofuginone and tryptophanol inhibited all of the above high glucose-induced alterations, indicating that activation of GCN-2 kinase ameliorates glucotoxicity in human peritoneal mesothelial cells, preserves their integrity, and prevents MMT. Whether such a strategy could be applied in the clinic to avoid ultrafiltration failure in PD patients remains to be investigated.

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