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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

Relative level of selected miRNAs in HEK293 cells following IGROV1 cell-secreted exosome treatment. qPCR was utilized to determine the relative levels and data were normalized against U6 snRNA. Asterisks indicate a p value <0.05.
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fig8: Relative level of selected miRNAs in HEK293 cells following IGROV1 cell-secreted exosome treatment. qPCR was utilized to determine the relative levels and data were normalized against U6 snRNA. Asterisks indicate a p value <0.05.

Mentions: In addition to EMT-related genes, relative changes in LIN28A, LIN28B, and selected miRNAs known to be involved in EMT and cancer (oncomirs) were assessed in HEK293 cells following uptake of IGROV1 cell-secreted exosomes. Taqman qPCR assays revealed that there was a significant, 15-fold increase in LIN28A level in HEK293 cells exposed to IGROV1 cell-secreted exosomes compared to IGROV1 cell conditioned media depleted of exosomes (Figure 6(a)), but not LIN28B (Figure 6(c)). This change was significant although no change in LIN28A (Figure 6(b)) or LIN28B (Figure 6(d)) protein level was evident in HEK293 cell following exposure to IGROV1 cell-secreted exosomes. No change in let-7 (a, b, c, d, ef, g, and i) miRNA was observed in HEK293 cells following treatment with IGROV1 cell-secreted exosomes (Figure 7). Moreover, qPCR analysis of selected other EMT and cancer associated miRNAs, that is, miR-17, miR-18a, miR-19a, miR-19b, miR-20a, miR-92, miR-22, miR-200a, miR-9, miR-30b, miR-30d, miR-30e, miR-125a-3p, miR-125-5p, and miR-125b, indicates a small but significant increase in miR-9 alone in HEK293 cells following treatment with IGROV1 cell-secreted exosomes (Figure 8).


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)

Relative level of selected miRNAs in HEK293 cells following IGROV1 cell-secreted exosome treatment. qPCR was utilized to determine the relative levels and data were normalized against U6 snRNA. Asterisks indicate a p value <0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4637063&req=5

fig8: Relative level of selected miRNAs in HEK293 cells following IGROV1 cell-secreted exosome treatment. qPCR was utilized to determine the relative levels and data were normalized against U6 snRNA. Asterisks indicate a p value <0.05.
Mentions: In addition to EMT-related genes, relative changes in LIN28A, LIN28B, and selected miRNAs known to be involved in EMT and cancer (oncomirs) were assessed in HEK293 cells following uptake of IGROV1 cell-secreted exosomes. Taqman qPCR assays revealed that there was a significant, 15-fold increase in LIN28A level in HEK293 cells exposed to IGROV1 cell-secreted exosomes compared to IGROV1 cell conditioned media depleted of exosomes (Figure 6(a)), but not LIN28B (Figure 6(c)). This change was significant although no change in LIN28A (Figure 6(b)) or LIN28B (Figure 6(d)) protein level was evident in HEK293 cell following exposure to IGROV1 cell-secreted exosomes. No change in let-7 (a, b, c, d, ef, g, and i) miRNA was observed in HEK293 cells following treatment with IGROV1 cell-secreted exosomes (Figure 7). Moreover, qPCR analysis of selected other EMT and cancer associated miRNAs, that is, miR-17, miR-18a, miR-19a, miR-19b, miR-20a, miR-92, miR-22, miR-200a, miR-9, miR-30b, miR-30d, miR-30e, miR-125a-3p, miR-125-5p, and miR-125b, indicates a small but significant increase in miR-9 alone in HEK293 cells following treatment with IGROV1 cell-secreted exosomes (Figure 8).

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