P2X7 promotes metastatic spreading and triggers release of miRNA-containing exosomes and microvesicles from melanoma cells

AbstractTumor growth and metastatic spreading are heavily affected by the P2X7 receptor as well as microvesicles and exosomes release into the tumor microenvironment. P2X7 receptor stimulation is known to trigger vesicular release from immune and central nervous system cells. However, P2X7 role in microvesicles and exosomes delivery from tumor cells was never analyzed in depth. Here we show that P2X7 is overexpressed in patients affected by metastatic malignant melanoma and that its expression closely correlates with reduced overall survival. Antagonism of melanoma cell-expressed P2X7 receptor inhibited in vitro anchorage-independent growth and migration and in vivo dissemination and lung metastasis formation. P2X7 stimulation triggered the release of miRNA-containing microvesicles and exosomes from melanoma cells, profoundly altering the nature of their miRNA content, as well as their dimensions and quantity. Among the more than 200 miRNAs that we found up-or-down-modulated for each vesicular fraction tested, we identified three miRNAs, miR-495-3p, miR-376c-3p, and miR-6730-3p, that were enriched in both the exosome and microvesicle fraction in a P2X7-dependent fashion. Interestingly, upon transfection, these miRNAs promoted melanoma cell growth or migration, and their vesicular release was minimized by P2X7 antagonism. Our data unveil an exosome/microvesicle and miRNA-dependent mechanism for the pro-metastatic activity of the P2X7 receptor and highlight this receptor as a suitable prognostic biomarker and therapeutic target in malignant melanoma.

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Luteolin inhibits proliferation and induces apoptosis of human melanoma cells in vivo and in vitro by suppressing MMP-2 and MMP-9 through the PI3K/AKT pathway

Food & Function ◽

10.1039/c8fo02013b ◽

2019 ◽

Vol 10 (2) ◽

pp. 703-712 ◽

Cited By ~ 19

Author(s):

Xin Yao ◽

Wei Jiang ◽

Danhong Yu ◽

Zhaowei Yan

Keyword(s):

Malignant Melanoma ◽

Incidence Rate ◽

Melanoma Cell ◽

Melanoma Cells ◽

Human Melanoma ◽

Akt Pathway ◽

Human Melanoma Cells ◽

Cell Metastasis

Since the incidence rate of malignant melanoma is increasing annually, development of drugs against melanoma cell metastasis has become more urgent.

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Functional Implication of Netrin Expression in Malignant Melanoma

Analytical Cellular Pathology ◽

10.1155/2009/954172 ◽

2009 ◽

Vol 31 (6) ◽

pp. 415-422

Author(s):

Simone Kaufmann ◽

Silke Kuphal ◽

Thomas Schubert ◽

Anja K. Bosserhoff

Keyword(s):

Malignant Melanoma ◽

Melanoma Cells ◽

Transcriptional Level ◽

Altered Expression ◽

Invasion And Migration ◽

Expression Of Genes ◽

Functional Relevance ◽

And Migration

Background: Malignant melanoma cells are known to have altered expression of genes supporting proliferation and invasion, however, the expression of molecules of the Netrin family of repellent factors has not been analyzed in melanomas until now.Results: Here, we show that Netrin-1 expression is strongly induced in melanoma cells compared to melanocytes in vivo and in vitro controlled at the transcriptional level via ETS-1. In addition, the expression of the netrin receptor UNC5B was induced and that of UNC5C was reduced in the tumor cells. In order to determine the functional relevance of Netrin-1 expression in malignant melanoma, Netrin expression in melanoma cells was reduced by siRNA attempts and primary human melanocytes were treated with recombinant Netrin-1. The cells showed no changes in proliferation or apoptosis, however, a strong reduction of migratory properties was observed in the melanoma cells after reduction of Netrin expression whereas melanocyte migration was strongly induced by treatment with Netrin.Conclusions: Our study suggests that Netrin-1 promotes melanoma cell invasion and migration and therefore has an important role in the progression of malignant melanoma.

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Total saponins from Rubus parvifolius L. inhibits cell proliferation, migration and invasion of malignant melanoma in vitro and in vivo

Bioscience Reports ◽

10.1042/bsr20201178 ◽

2020 ◽

Author(s):

Jinfeng Cao ◽

Xue Zhao ◽

Yan Ma ◽

Jian Yang ◽

Fuqiang Li

Keyword(s):

Cell Proliferation ◽

Malignant Melanoma ◽

Melanoma Cell ◽

Nude Mice ◽

Melanoma Metastasis ◽

Invasion And Migration ◽

Rubus Parvifolius ◽

And Migration

Background: Total saponins from Rubus parvifolius L. (TSRP) is the main bioactive fractions responsible for the antitumor activities. The work was aimed to evaluate the anti-tumor effect of TSRP in malignant melanoma in vitro and in vivo. Methods and Results: Anti melanoma cell proliferation, invasion and migration effect of TSRP were detected in human malignant melanoma A375 cells under the indicated time and dosages. In vivo anti-tumor effect of TSRP was measured in A375 xenograft immunodeficient nude mice. Sixty A375 xenografts were randomly divided into five groups: Vehicle, cyclophosphamide (CTX, 20 mg/kg), TSRP (25 mg/kg), TSRP (50 mg/kg) and TSRP (100 mg/kg) groups for 14 days’ treatment. In addition, the melanoma cell metastasis in lung in vivo of TSRP was detected in A375 tail vein injection mice, and the histopathalogical analysis of the metastasis lung was detected by H & E stating. TSRP was significantly inhibited the cell proliferation, invasion and migration of A375 in vitro at the indicated time and dosages. TSRP treatment was effectively blocked the tumor growth in immunodeficient nude mice. In addition, TSRP was also significantly inhibited the melanoma metastasis of lung. Conclusion: This study indicated that the TSRP has a remarkable anti malignant melanoma effect, which mainly through the inhibition of the cell invasion,migration and tumor metastasis.

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Isorhamnetin Induces Melanoma Cell Apoptosis via the PI3K/Akt and NF-κB Pathways

BioMed Research International ◽

10.1155/2020/1057943 ◽

2020 ◽

Vol 2020 ◽

pp. 1-11 ◽

Cited By ~ 1

Author(s):

Ran Duan ◽

Xiao Liang ◽

Bangda Chai ◽

Yiwen Zhou ◽

Hengyu Du ◽

...

Keyword(s):

Malignant Melanoma ◽

Melanoma Cell ◽

Cell Apoptosis ◽

In Vitro Study ◽

B16f10 Cell ◽

Antitumor Effects ◽

Melanoma Treatment ◽

And Migration

Malignant melanoma is characterized by its bad prognosis for aggressiveness, drug resistance, and early metastasis. Isorhamnetin (3′-methoxy-3,4′,5,7-tetrahydroxyflavone; IH) is a natural flavonoid that has been investigated for its antitumor effects in breast cancer, colon cancer, and gastric cancer through inducing cell apoptosis. Given its role in tumor inhibition, no research has been conducted concerning its effect against melanoma. In the present study, we found that IH could significantly inhibit B16F10 cell proliferation and migration and induce B16F10 cell apoptosis. The examination on molecular mechanism revealed that IH could suppress the phosphorylation of Akt and the translocation of NF-κB, which are key factors in apoptosis-related pathways. We also detected that this process was related to the bifunctional 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatases 4 (PFKFB4) by PFKFB4 knockdown experiment. In line with in vitro study, we further provided that IH effectively inhibited tumor growth in vivo. Taken together, IH was proven to induce melanoma cell apoptosis in vitro and in vivo, which may serve as a potential agent in malignant melanoma treatment in the future.

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Chondroitin polymerizing factor (CHPF) promotes development of malignant melanoma through regulation of CDK1

Cell Death and Disease ◽

10.1038/s41419-020-2526-9 ◽

2020 ◽

Vol 11 (7) ◽

Author(s):

Wei Sun ◽

Fang Zhao ◽

Yu Xu ◽

Kai Huang ◽

Xianling Guo ◽

...

Keyword(s):

Malignant Melanoma ◽

Pathway Analysis ◽

Normal Skin ◽

Over Expression ◽

Tumor Promotor ◽

And Migration ◽

First Time ◽

The Relationship

Abstract Chondroitin polymerizing factor (CHPF) is an important member of glycosyltransferases involved in the biosynthesis of chondroitin sulfate (CS). However, the relationship between CHPF and malignant melanoma (MM) is still unknown. In this study, it was demonstrated that CHPF was up-regulated in MM tissues compared with the adjacent normal skin tissues and its high expression was correlated with more advanced T stage. Further investigations indicated that the over-expression/knockdown of CHPF could promote/inhibit proliferation, colony formation and migration of MM cells, while inhibiting/promoting cell apoptosis. Moreover, knockdown of CHPF could also suppress tumorigenicity of MM cells in vivo. RNA-sequencing followed by Ingenuity pathway analysis (IPA) was performed for exploring downstream of CHPF and identified CDK1 as the potential target. Furthermore, our study revealed that knockdown of CDK1 could inhibit development of MM in vitro, and alleviate the CHPF over-expression induced promotion of MM. In conclusion, our study showed, as the first time, CHPF as a tumor promotor for MM, whose function was carried out probably through the regulation of CDK1.

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Treatment of melanoma cells with the synthetic retinoid CD437 induces apoptosis via activation of AP-1 in vitro, and causes growth inhibition in xenografts in vivo.

The Journal of Cell Biology ◽

10.1083/jcb.135.6.1889 ◽

1996 ◽

Vol 135 (6) ◽

pp. 1889-1898 ◽

Cited By ~ 75

Author(s):

D Schadendorf ◽

M A Kern ◽

M Artuc ◽

H L Pahl ◽

T Rosenbach ◽

...

Keyword(s):

Cell Death ◽

Malignant Melanoma ◽

Programmed Cell Death ◽

Melanoma Cells ◽

Human Melanoma ◽

Mrna Levels ◽

Synthetic Retinoid ◽

Human Melanoma Cells

Human malignant melanoma is notoriously resistant to pharmacological modulation. We describe here for the first time that the synthetic retinoid CD437 has a strong dose-dependent antiproliferative effect on human melanoma cells (IC50: 5 x 10(-6) M) via the induction of programmed cell death, as judged by analysis of cell morphology, electron microscopical features, and DNA fragmentation. Programmed cell death was preceded by a strong activation of the AP-1 complex in CD437-treated cells as demonstrated by gel retardation and chloramphenicol transferase (CAT) assays. Northern blot analysis showed a time-dependent increase in the expression of c-fos and c-jun encoding components of AP-1, whereas bcl-2 and p53 mRNA levels remained constant. CD437 also exhibited a strong growth inhibitory effect on MeWo melanoma cells in a xenograft model. In tissue sections of CD437-treated MeWo tumors from these animals, apoptotic melanoma cells and c-fos overexpressing cells were colocalized by TdT-mediated deoxyuridine triphosphate-digoxigenin nick end labeling (TUNEL) staining and in situ hybridization. Taken together, this report identifies CD437 as a retinoid that activates and upregulates the transcription factor AP-1, leading eventually to programmed cell death of exposed human melanoma cells in vitro and in vivo. Further studies are needed to evaluate whether synthetic retinoids such as CD437 represent a new class of retinoids, which may open up new ways to a more effective therapy of malignant melanoma.

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Tumour Progression Stage-Dependent Secretion of YB-1 Stimulates Melanoma Cell Migration and Invasion

Cancers ◽

10.3390/cancers12082328 ◽

2020 ◽

Vol 12 (8) ◽

pp. 2328 ◽

Cited By ~ 2

Author(s):

Corinna Kosnopfel ◽

Tobias Sinnberg ◽

Birgit Sauer ◽

Heike Niessner ◽

Alina Muenchow ◽

...

Keyword(s):

Cell Migration ◽

Malignant Melanoma ◽

Melanoma Cell ◽

Cell Biology ◽

Secretory Pathway ◽

Tumour Progression ◽

Melanoma Cells ◽

Migration And Invasion ◽

Melanoma Progression ◽

And Migration

Secreted factors play an important role in intercellular communication. Therefore, they are not only indispensable for the regulation of various physiological processes but can also decisively advance the development and progression of tumours. In the context of inflammatory disease, Y-box binding protein 1 (YB-1) is actively secreted and the extracellular protein promotes cell proliferation and migration. In malignant melanoma, intracellular YB-1 expression increases during melanoma progression and represents an unfavourable prognostic marker. Here, we show active secretion of YB-1 from melanoma cells as opposed to benign cells of the skin. Intriguingly, YB-1 secretion correlates with the stage of melanoma progression and depends on a calcium- and ATP-dependent non-classical secretory pathway leading to the occurrence of YB-1 in the extracellular space as a free protein. Along with an elevated YB-1 secretion of melanoma cells in the metastatic growth phase, extracellular YB-1 exerts a stimulating effect on melanoma cell migration, invasion, and tumourigenicity. Collectively, these data suggest that secreted YB-1 plays a functional role in melanoma cell biology, stimulating metastasis, and may serve as a novel biomarker in malignant melanoma that reflects tumour aggressiveness.

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Suppression of long noncoding RNA MALAT1 inhibits the development of uveal melanoma via microRNA-608-mediated inhibition of HOXC4

AJP Cell Physiology ◽

10.1152/ajpcell.00262.2019 ◽

2020 ◽

Vol 318 (5) ◽

pp. C903-C912 ◽

Cited By ~ 3

Author(s):

Shuai Wu ◽

Han Chen ◽

Ling Zuo ◽

Hai Jiang ◽

Hongtao Yan

Keyword(s):

Cell Cycle ◽

Uveal Melanoma ◽

Noncoding Rna ◽

Cell Cycle Progression ◽

Melanoma Cells ◽

Cell Cycle Distribution ◽

Invasion And Migration ◽

And Migration

This study explored the effects of the metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) on the development of uveal melanoma. Moreover, the role of the MALAT1/microRNA-608 (miR-608)/homeobox C4 (HOXC4) axis was assessed by evaluating the proliferation, invasion, and migration, as well as the cell cycle distribution of uveal melanoma in vitro after knocking down MALAT1 or HOXC4 and/or overexpression of miR-608 in uveal melanoma cells (MUM-2B and C918). Moreover, the effects of the MALAT1/miR-608/HOXC4 axis in uveal melanoma in vivo were further evaluated by injecting the C918 cells into the NOD/SCID mice. HOXC4 was found to be a gene upregulated in uveal melanoma, while knockdown of its expression resulted in suppression of uveal melanoma cell migration, proliferation, and invasion, as well as cell cycle progression. In addition, the upregulation of miR-608 reduced the expression of HOXC4 in the uveal melanoma cells, which was rescued by overexpression of MALAT1. Hence, MALAT1 could upregulate the HOXC4 by binding to miR-608. The suppressed progression of uveal melanoma in vitro by miR-608 was rescued by overexpression of MALAT1. Additionally, in vivo assays demonstrated that downregulation of MALAT1 could suppress tumor growth through downregulation of HOXC4 expression via increasing miR-608 in uveal melanoma. In summary, MALAT1 downregulation functions to restrain the development of uveal melanoma via miR-608-mediated inhibition of HOXC4.

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Capsaicin Exerts Therapeutic Effects by targeting tNOX-SIRT1 Axis and Augmenting ROS-Dependent Cytotoxic Autophagy in Melanoma Cancer Cells

10.21203/rs.3.rs-117713/v1 ◽

2020 ◽

Author(s):

Atikul Islam ◽

Pei-Fang Hsieh ◽

Jou-Chun Chou ◽

Jiunn-Wang Liao ◽

Ming-Kun Hsieh ◽

...

Keyword(s):

Malignant Melanoma ◽

Cancer Cells ◽

Nadh Oxidase ◽

Treatment Options ◽

Melanoma Cells ◽

Therapeutic Effects ◽

Melanoma Cancer ◽

Anticancer Properties

Abstract Background: Although considered a rare form of skin cancer, malignant melanoma has steadily increased internationally and is a main cause of cancer-associated death worldwide. The treatment options for malignant melanoma are very limited. Accumulating data suggest that the natural compound, capsaicin, exhibits preferential anticancer properties to act as a nutraceutical agent. Here, we explored the underlying molecular events involved in the inhibitory effects of capsaicin on the growth of melanoma cells.Methods: The cellular thermal shift assay (CETSA) and isothermal dose response fingerprint (ITDRFCETSA) were utilized to validate the binding of capsaicin with the tumor-associated NADH oxidase, tNOX (ENOX2) in melanoma cells. We also assessed the cellular impact of capsaicin-targeting of tNOX on A375 cells by flow cytometry and protein analysis. The essential role of tNOX in tumor- and melanoma-growth limiting abilities of capsaicin was evaluated in C57BL/6 mice.Results: Our data show that capsaicin directly targets cellular tNOX to inhibit its enzymatic activity and enhance protein degradation capacity. The inhibition of tNOX by capsaicin is accompanied by the attenuation of SIRT1, a NAD+-dependent deacetylase that enhances ULK1 acetylation to induce ROS-dependent autophagy in melanoma cells. Capsaicin treatment of mice implanted with melanoma cancer cells suppressed tumor growth by down-regulating tNOX and SIRT1, which was also seen in an in vivo xenograft study with tNOX-depleted melanoma cells. Conclusions: Together, our findings suggest that tNOX expression is important for the growth of melanoma cancer cells both in vitro and in vivo, and that inhibition of the tNOX-SIRT1 axis contributes to inducting cytotoxic ROS-dependent autophagy in melanoma cells.

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