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High LIN28A Expressing Ovarian Cancer Cells Secrete Exosomes That Induce Invasion and Migration in HEK293 Cells.

Enriquez VA, Cleys ER, Da Silveira JC, Spillman MA, Winger QA, Bouma GJ - Biomed Res Int (2015)

Bottom Line: IGROV1 cells were found to contain high LIN28A and secrete exosomes that were taken up by HEK293 cells.Moreover, exposure to these IGROV1 secreted exosomes led to significant increases in genes involved in Epithelial-to-Mesenchymal Transition (EMT), induced HEK293 cell invasion and migration.No evidence was found of LIN28A transfer from IGROV1 exosomes to HEK293 cells.

View Article: PubMed Central - PubMed

Affiliation: Cell and Molecular Biology Program, Department of Biomedical Sciences, Colorado State University, Fort Collins, CO 80523-1683, USA ; Animal Reproduction and Biotechnology Laboratory, Department of Biomedical Sciences, Colorado State University, Fort Collins, CO 80523-1683, USA.

ABSTRACT
Epithelial ovarian cancer is the most aggressive and deadly form of ovarian cancer and is the most lethal gynecological malignancy worldwide; therefore, efforts to elucidate the molecular factors that lead to epithelial ovarian cancer are essential to better understand this disease. Recent studies reveal that tumor cells release cell-secreted vesicles called exosomes and these exosomes can transfer RNAs and miRNAs to distant sites, leading to cell transformation and tumor development. The RNA-binding protein LIN28 is a known marker of stem cells and when expressed in cancer, it is associated with poor tumor outcome. We hypothesized that high LIN28 expressing ovarian cancer cells secrete exosomes that can be taken up by nontumor cells and cause changes in gene expression and cell behavior associated with tumor development. IGROV1 cells were found to contain high LIN28A and secrete exosomes that were taken up by HEK293 cells. Moreover, exposure to these IGROV1 secreted exosomes led to significant increases in genes involved in Epithelial-to-Mesenchymal Transition (EMT), induced HEK293 cell invasion and migration. These changes were not observed with exosomes secreted by OV420 cells, which contain no detectable amounts of LIN28A or LIN28B. No evidence was found of LIN28A transfer from IGROV1 exosomes to HEK293 cells.

No MeSH data available.


Related in: MedlinePlus

Detection of exosome-uptake by HEK293 cells. (a) HEK293 cells after exposure to CD63-GFP positive (green) and DiD-RFP labeled (red) IGROV1 secreted exosomes. 20x magnification was utilized to image HEK293 cells following exosome exposure using confocal microscopy. (b) Z-stack image of HEK293 cells after exposure to CD63-GFP positive (green) and DiD-RFP labeled (red) IGROV1 secreted exosomes. (c) Z-stack image of HEK293 cells after exposure to OV420 secreted, DiD labeled (red) exosomes. 40x magnification was utilized to image HEK293 cells following exosome exposure using confocal microscopy.
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fig3: Detection of exosome-uptake by HEK293 cells. (a) HEK293 cells after exposure to CD63-GFP positive (green) and DiD-RFP labeled (red) IGROV1 secreted exosomes. 20x magnification was utilized to image HEK293 cells following exosome exposure using confocal microscopy. (b) Z-stack image of HEK293 cells after exposure to CD63-GFP positive (green) and DiD-RFP labeled (red) IGROV1 secreted exosomes. (c) Z-stack image of HEK293 cells after exposure to OV420 secreted, DiD labeled (red) exosomes. 40x magnification was utilized to image HEK293 cells following exosome exposure using confocal microscopy.

Mentions: IGROV1 cells were infected with CD63-GFP-cytotracer, which led to GFP-labeling of exosomes by the host cells. In addition, exosomes secreted by OV420 cells were incubated with the DiD cell-labeling solution, which labels lipids in cell membranes. HEK293 cell incubation with GFP-labeled IGROV1 secreted exosomes or DiD-labeled OV420 secreted exosomes leads to uptake of these exosomes as evident by Z-stack imaging (Figure 3).


High LIN28A Expressing Ovarian Cancer Cells Secrete Exosomes That Induce Invasion and Migration in HEK293 Cells.

Enriquez VA, Cleys ER, Da Silveira JC, Spillman MA, Winger QA, Bouma GJ - Biomed Res Int (2015)

Detection of exosome-uptake by HEK293 cells. (a) HEK293 cells after exposure to CD63-GFP positive (green) and DiD-RFP labeled (red) IGROV1 secreted exosomes. 20x magnification was utilized to image HEK293 cells following exosome exposure using confocal microscopy. (b) Z-stack image of HEK293 cells after exposure to CD63-GFP positive (green) and DiD-RFP labeled (red) IGROV1 secreted exosomes. (c) Z-stack image of HEK293 cells after exposure to OV420 secreted, DiD labeled (red) exosomes. 40x magnification was utilized to image HEK293 cells following exosome exposure using confocal microscopy.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4637063&req=5

fig3: Detection of exosome-uptake by HEK293 cells. (a) HEK293 cells after exposure to CD63-GFP positive (green) and DiD-RFP labeled (red) IGROV1 secreted exosomes. 20x magnification was utilized to image HEK293 cells following exosome exposure using confocal microscopy. (b) Z-stack image of HEK293 cells after exposure to CD63-GFP positive (green) and DiD-RFP labeled (red) IGROV1 secreted exosomes. (c) Z-stack image of HEK293 cells after exposure to OV420 secreted, DiD labeled (red) exosomes. 40x magnification was utilized to image HEK293 cells following exosome exposure using confocal microscopy.
Mentions: IGROV1 cells were infected with CD63-GFP-cytotracer, which led to GFP-labeling of exosomes by the host cells. In addition, exosomes secreted by OV420 cells were incubated with the DiD cell-labeling solution, which labels lipids in cell membranes. HEK293 cell incubation with GFP-labeled IGROV1 secreted exosomes or DiD-labeled OV420 secreted exosomes leads to uptake of these exosomes as evident by Z-stack imaging (Figure 3).

Bottom Line: IGROV1 cells were found to contain high LIN28A and secrete exosomes that were taken up by HEK293 cells.Moreover, exposure to these IGROV1 secreted exosomes led to significant increases in genes involved in Epithelial-to-Mesenchymal Transition (EMT), induced HEK293 cell invasion and migration.No evidence was found of LIN28A transfer from IGROV1 exosomes to HEK293 cells.

View Article: PubMed Central - PubMed

Affiliation: Cell and Molecular Biology Program, Department of Biomedical Sciences, Colorado State University, Fort Collins, CO 80523-1683, USA ; Animal Reproduction and Biotechnology Laboratory, Department of Biomedical Sciences, Colorado State University, Fort Collins, CO 80523-1683, USA.

ABSTRACT
Epithelial ovarian cancer is the most aggressive and deadly form of ovarian cancer and is the most lethal gynecological malignancy worldwide; therefore, efforts to elucidate the molecular factors that lead to epithelial ovarian cancer are essential to better understand this disease. Recent studies reveal that tumor cells release cell-secreted vesicles called exosomes and these exosomes can transfer RNAs and miRNAs to distant sites, leading to cell transformation and tumor development. The RNA-binding protein LIN28 is a known marker of stem cells and when expressed in cancer, it is associated with poor tumor outcome. We hypothesized that high LIN28 expressing ovarian cancer cells secrete exosomes that can be taken up by nontumor cells and cause changes in gene expression and cell behavior associated with tumor development. IGROV1 cells were found to contain high LIN28A and secrete exosomes that were taken up by HEK293 cells. Moreover, exposure to these IGROV1 secreted exosomes led to significant increases in genes involved in Epithelial-to-Mesenchymal Transition (EMT), induced HEK293 cell invasion and migration. These changes were not observed with exosomes secreted by OV420 cells, which contain no detectable amounts of LIN28A or LIN28B. No evidence was found of LIN28A transfer from IGROV1 exosomes to HEK293 cells.

No MeSH data available.


Related in: MedlinePlus