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Extracellular vesicle-mediated phenotype switching in malignant and non-malignant colon cells.

Mulvey HE, Chang A, Adler J, Del Tatto M, Perez K, Quesenberry PJ, Chatterjee D - BMC Cancer (2015)

Bottom Line: We also demonstrate that knock down of 14-3-3 zeta/delta reduced anchorage-independent growth of HCT116 cells and 1459 cells co-cultured with HCT derived EVs.Evidence of EV-mediated induction of malignant phenotype, and reversal of malignant phenotype, provides rational basis for further study of the role of EVs in tumorigenesis.Identification of 14-3-3 zeta/delta as up-regulated in malignancy suggests its potential as a putative drug target for the treatment of colorectal cancer.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Rhode Island Hospital and The Alpert Medical School of Brown University, Coro West, Suite 5.01, One Hoppin St, Providence, RI, 02903, USA. hillary.e.mulvey@gmail.com.

ABSTRACT

Background: Extracellular vesicles (EVs) are secreted from many cells, carrying cargoes including proteins and nucleic acids. Research has shown that EVs play a role in a variety of biological processes including immunity, bone formation and recently they have been implicated in promotion of a metastatic phenotype.

Methods: EVs were isolated from HCT116 colon cancer cells, 1459 non-malignant colon fibroblast cells, and tumor and normal colon tissue from a patient sample. Co-cultures were performed with 1459 cells and malignant vesicles, as well as HCT116 cells and non-malignant vesicles. Malignant phenotype was measured using soft agar colony formation assay. Co-cultures were also analyzed for protein levels using mass spectrometry. The importance of 14-3-3 zeta/delta in transfer of malignant phenotype was explored using siRNA. Additionally, luciferase reporter assay was used to measure the transcriptional activity of NF-κB.

Results: This study demonstrates the ability of EVs derived from malignant colon cancer cell line and malignant patient tissue to induce the malignant phenotype in non-malignant colon cells. Similarly, EVs derived from non-malignant colon cell lines and normal patient tissue reversed the malignant phenotype of HCT116 cells. Cells expressing an EV-induced malignant phenotype showed increased transcriptional activity of NF-κB which was inhibited by the NF--κB inhibitor, BAY117082. We also demonstrate that knock down of 14-3-3 zeta/delta reduced anchorage-independent growth of HCT116 cells and 1459 cells co-cultured with HCT derived EVs.

Conclusions: Evidence of EV-mediated induction of malignant phenotype, and reversal of malignant phenotype, provides rational basis for further study of the role of EVs in tumorigenesis. Identification of 14-3-3 zeta/delta as up-regulated in malignancy suggests its potential as a putative drug target for the treatment of colorectal cancer.

No MeSH data available.


Related in: MedlinePlus

Schematic representing our model indicating the transfer of 14-3-3 zeta/delta (circles) by HCT116 EVs with the activation of NF-κB with downstream signaling events regulated by both 14-3-3 zeta/delta and NF-κB leading to the malignant phenotype that is inhibited by non-malignant EVs
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Fig6: Schematic representing our model indicating the transfer of 14-3-3 zeta/delta (circles) by HCT116 EVs with the activation of NF-κB with downstream signaling events regulated by both 14-3-3 zeta/delta and NF-κB leading to the malignant phenotype that is inhibited by non-malignant EVs

Mentions: Collectively, our results indicate that HCT116 EVs carry 14-3-3 zeta/delta as part of their cargo that is transferred to recipient cells after co-culture (Fig. 6). This also results in the activation of NF-κB (Fig. 6). Consequently, there is a promotion of a series of events leading to HCT116 cancer cell survival, which can be abrogated, in part, by co-culture with non-malignant EVs (Fig. 6).Fig. 6


Extracellular vesicle-mediated phenotype switching in malignant and non-malignant colon cells.

Mulvey HE, Chang A, Adler J, Del Tatto M, Perez K, Quesenberry PJ, Chatterjee D - BMC Cancer (2015)

Schematic representing our model indicating the transfer of 14-3-3 zeta/delta (circles) by HCT116 EVs with the activation of NF-κB with downstream signaling events regulated by both 14-3-3 zeta/delta and NF-κB leading to the malignant phenotype that is inhibited by non-malignant EVs
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4522096&req=5

Fig6: Schematic representing our model indicating the transfer of 14-3-3 zeta/delta (circles) by HCT116 EVs with the activation of NF-κB with downstream signaling events regulated by both 14-3-3 zeta/delta and NF-κB leading to the malignant phenotype that is inhibited by non-malignant EVs
Mentions: Collectively, our results indicate that HCT116 EVs carry 14-3-3 zeta/delta as part of their cargo that is transferred to recipient cells after co-culture (Fig. 6). This also results in the activation of NF-κB (Fig. 6). Consequently, there is a promotion of a series of events leading to HCT116 cancer cell survival, which can be abrogated, in part, by co-culture with non-malignant EVs (Fig. 6).Fig. 6

Bottom Line: We also demonstrate that knock down of 14-3-3 zeta/delta reduced anchorage-independent growth of HCT116 cells and 1459 cells co-cultured with HCT derived EVs.Evidence of EV-mediated induction of malignant phenotype, and reversal of malignant phenotype, provides rational basis for further study of the role of EVs in tumorigenesis.Identification of 14-3-3 zeta/delta as up-regulated in malignancy suggests its potential as a putative drug target for the treatment of colorectal cancer.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Rhode Island Hospital and The Alpert Medical School of Brown University, Coro West, Suite 5.01, One Hoppin St, Providence, RI, 02903, USA. hillary.e.mulvey@gmail.com.

ABSTRACT

Background: Extracellular vesicles (EVs) are secreted from many cells, carrying cargoes including proteins and nucleic acids. Research has shown that EVs play a role in a variety of biological processes including immunity, bone formation and recently they have been implicated in promotion of a metastatic phenotype.

Methods: EVs were isolated from HCT116 colon cancer cells, 1459 non-malignant colon fibroblast cells, and tumor and normal colon tissue from a patient sample. Co-cultures were performed with 1459 cells and malignant vesicles, as well as HCT116 cells and non-malignant vesicles. Malignant phenotype was measured using soft agar colony formation assay. Co-cultures were also analyzed for protein levels using mass spectrometry. The importance of 14-3-3 zeta/delta in transfer of malignant phenotype was explored using siRNA. Additionally, luciferase reporter assay was used to measure the transcriptional activity of NF-κB.

Results: This study demonstrates the ability of EVs derived from malignant colon cancer cell line and malignant patient tissue to induce the malignant phenotype in non-malignant colon cells. Similarly, EVs derived from non-malignant colon cell lines and normal patient tissue reversed the malignant phenotype of HCT116 cells. Cells expressing an EV-induced malignant phenotype showed increased transcriptional activity of NF-κB which was inhibited by the NF--κB inhibitor, BAY117082. We also demonstrate that knock down of 14-3-3 zeta/delta reduced anchorage-independent growth of HCT116 cells and 1459 cells co-cultured with HCT derived EVs.

Conclusions: Evidence of EV-mediated induction of malignant phenotype, and reversal of malignant phenotype, provides rational basis for further study of the role of EVs in tumorigenesis. Identification of 14-3-3 zeta/delta as up-regulated in malignancy suggests its potential as a putative drug target for the treatment of colorectal cancer.

No MeSH data available.


Related in: MedlinePlus