Role of the Exosome Secretion Machinery in Ovarian Carcinoma: In Vitro and In Vivo Models
Joint Authors
Onallah, Hadil
Davidson, Ben
Reich, Reuven
Broner, Esther Channah
Tavor Re’em, Tali
Source
Issue
Vol. 2020, Issue 2020 (31 Dec. 2020), pp.1-11, 11 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2020-05-30
Country of Publication
Egypt
No. of Pages
11
Main Subjects
Abstract EN
Objective.
We recently reported on the expression and clinical role of molecules that mediate exosome secretion in high-grade serous carcinoma.
In the present study, the biological role of these molecules was analyzed.
Methods.
OVCAR8 and ES-2 ovarian carcinoma cells were studied using a combination of CRISPR/Cas9 technology and two 3D in vitro models—spheroids emulating effusions and alginate scaffolds representing solid lesions.
Isolation of exosomes was validated by electron microscopy.
TSAP6, NSMASE2, RAB27A, and RAB27B mRNA and protein levels were analyzed using qRT-PCR and Western blotting, respectively.
Tumor aggressiveness was studied in vitro using scratch assay, invasion assay, and matrix metalloproteinase (MMP) activity assay and in vivo using a mouse model.
Results.
In OVCAR8 cells, mRNA expression of TSAP6 and RAB27A was significantly higher in spheroids compared to scaffolds, whereas the opposite was true for NSMASE2 mRNA.
In ES-2 cells, TSAP6 and RAB27B mRNA expression was significantly higher in spheroids versus scaffolds.
In addition, nSMase2 and TSAP6 protein expression was significantly higher in scaffolds compared to spheroids.
CRISPR-edited cells with silencing of NSMASE2, TSAP6, and RAB27A/B had reduced migration, invasion, and MMP activity.
Additionally, knockout (KO) of these molecules resulted in significantly diminished exosome secretion.
In vivo assay showed that when injected to mice, OVCAR8 RAB27A/B KO cells, as opposed to control OVCAR8 cells, did not form ascites or visible tumor lesions and had reduced MMP expression.
Conclusion.
The present study provides evidence that different models for culturing ovarian carcinoma cells affect the expression of molecules mediating exosome secretion and that these molecules have a tumor-promoting role.
Silencing these molecules may consequently have therapeutic relevance in this cancer.
American Psychological Association (APA)
Broner, Esther Channah& Onallah, Hadil& Tavor Re’em, Tali& Davidson, Ben& Reich, Reuven. 2020. Role of the Exosome Secretion Machinery in Ovarian Carcinoma: In Vitro and In Vivo Models. Journal of Oncology،Vol. 2020, no. 2020, pp.1-11.
https://search.emarefa.net/detail/BIM-1188948
Modern Language Association (MLA)
Broner, Esther Channah…[et al.]. Role of the Exosome Secretion Machinery in Ovarian Carcinoma: In Vitro and In Vivo Models. Journal of Oncology No. 2020 (2020), pp.1-11.
https://search.emarefa.net/detail/BIM-1188948
American Medical Association (AMA)
Broner, Esther Channah& Onallah, Hadil& Tavor Re’em, Tali& Davidson, Ben& Reich, Reuven. Role of the Exosome Secretion Machinery in Ovarian Carcinoma: In Vitro and In Vivo Models. Journal of Oncology. 2020. Vol. 2020, no. 2020, pp.1-11.
https://search.emarefa.net/detail/BIM-1188948
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1188948