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Endothelial exosomes contribute to the antitumor response during breast cancer neoadjuvant chemotherapy via microRNA transfer.

Bovy N, Blomme B, Frères P, Dederen S, Nivelles O, Lion M, Carnet O, Martial JA, Noël A, Thiry M, Jérusalem G, Josse C, Bours V, Tabruyn SP, Struman I - Oncotarget (2015)

Bottom Line: The modulation of miR-503 in breast cancer cells altered their proliferative and invasive capacities.We then identified two targets of miR-503, CCND2 and CCND3.Moreover, we measured increased plasmatic miR-503 in breast cancer patients after neoadjuvant chemotherapy, which could be partly due to increased miRNA secretion by endothelial cells.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Molecular Angiogenesis, GIGA-R, University of Liège, Belgium.

ABSTRACT
The interaction between tumor cells and their microenvironment is an essential aspect of tumor development. Therefore, understanding how this microenvironment communicates with tumor cells is crucial for the development of new anti-cancer therapies. MicroRNAs (miRNAs) are small non-coding RNAs that inhibit gene expression. They are secreted into the extracellular medium in vesicles called exosomes, which allow communication between cells via the transfer of their cargo. Consequently, we hypothesized that circulating endothelial miRNAs could be transferred to tumor cells and modify their phenotype. Using exogenous miRNA, we demonstrated that endothelial cells can transfer miRNA to tumor cells via exosomes. Using miRNA profiling, we identified miR-503, which exhibited downregulated levels in exosomes released from endothelial cells cultured under tumoral conditions. The modulation of miR-503 in breast cancer cells altered their proliferative and invasive capacities. We then identified two targets of miR-503, CCND2 and CCND3. Moreover, we measured increased plasmatic miR-503 in breast cancer patients after neoadjuvant chemotherapy, which could be partly due to increased miRNA secretion by endothelial cells. Taken together, our data are the first to reveal the involvement of the endothelium in the modulation of tumor development via the secretion of circulating miR-503 in response to chemotherapy treatment.

No MeSH data available.


Related in: MedlinePlus

Endothelial miR-503 impairs tumor growth in vitro(A) Invasion level of MDA-MB-231 cells transfected with pre-miR-control or pre-miR-503 and with anti-miR-control or anti-miR-503. (B) Luminescence quantification of MDA-MB-231 cells transfected with pre-miR-control or pre-miR-503 and with anti-miR-control or anti-miR-503. (C) MiR-503 levels, measured by qRT-PCR in MDA-MB-231 cells incubated with 5 μg of HUVEC exosomes for 24 h. (D) Invasion level of MDA-MB-231 cells transfected with anti-miR-control or anti-miR-503 and cocultured with HUVECs transfected with pre-miR-control or pre-miR-503. (E) Luminescence quantification of MDA-MB-231 cells transfected with anti-miR-control or anti-miR-503 and cocultured with HUVECs transfected with pre-miR-control or pre-miR-503. (F) Luminescence quantification of MDA-MB-231 cells incubated with exosomes from HUVECs transfected with pre-miR-control or pre-miR-503. Additionally, see Fig. S3.
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Figure 3: Endothelial miR-503 impairs tumor growth in vitro(A) Invasion level of MDA-MB-231 cells transfected with pre-miR-control or pre-miR-503 and with anti-miR-control or anti-miR-503. (B) Luminescence quantification of MDA-MB-231 cells transfected with pre-miR-control or pre-miR-503 and with anti-miR-control or anti-miR-503. (C) MiR-503 levels, measured by qRT-PCR in MDA-MB-231 cells incubated with 5 μg of HUVEC exosomes for 24 h. (D) Invasion level of MDA-MB-231 cells transfected with anti-miR-control or anti-miR-503 and cocultured with HUVECs transfected with pre-miR-control or pre-miR-503. (E) Luminescence quantification of MDA-MB-231 cells transfected with anti-miR-control or anti-miR-503 and cocultured with HUVECs transfected with pre-miR-control or pre-miR-503. (F) Luminescence quantification of MDA-MB-231 cells incubated with exosomes from HUVECs transfected with pre-miR-control or pre-miR-503. Additionally, see Fig. S3.

Mentions: To investigate the impact of miR-503 on tumor growth, we performed gain- and loss-of-function studies. MDA-MB-231 cells were transfected with either pre- or anti-miR-503, and the transfection efficiency was monitored using qRT-PCR (Fig. S3A). We used MDA-MB-231 constitutively expressing luciferase to quantify the proliferation in a coculture system by measuring the luminescence. Moreover, tumor cell invasion was assessed by quantifying the sprouting of tumor spheroids in a 3D collagen matrix. Increasing miR-503 levels via the transfection of miRNA mimics (pre-miRs) in MDA-MB-231 cells decreased both cell proliferation and invasion. Conversely, inhibition of miR-503 via the transfection of miR-503 antisense LNAs (anti-miRs) resulted in increased levels both of these processes compared with those of the control (Fig. 3A-B). Moreover, the effects of modulating miR-503 on tumor proliferation and invasion were confirmed by measuring BrdU incorporation and performing Boyden chamber assays, respectively (Fig. S3B-C).


Endothelial exosomes contribute to the antitumor response during breast cancer neoadjuvant chemotherapy via microRNA transfer.

Bovy N, Blomme B, Frères P, Dederen S, Nivelles O, Lion M, Carnet O, Martial JA, Noël A, Thiry M, Jérusalem G, Josse C, Bours V, Tabruyn SP, Struman I - Oncotarget (2015)

Endothelial miR-503 impairs tumor growth in vitro(A) Invasion level of MDA-MB-231 cells transfected with pre-miR-control or pre-miR-503 and with anti-miR-control or anti-miR-503. (B) Luminescence quantification of MDA-MB-231 cells transfected with pre-miR-control or pre-miR-503 and with anti-miR-control or anti-miR-503. (C) MiR-503 levels, measured by qRT-PCR in MDA-MB-231 cells incubated with 5 μg of HUVEC exosomes for 24 h. (D) Invasion level of MDA-MB-231 cells transfected with anti-miR-control or anti-miR-503 and cocultured with HUVECs transfected with pre-miR-control or pre-miR-503. (E) Luminescence quantification of MDA-MB-231 cells transfected with anti-miR-control or anti-miR-503 and cocultured with HUVECs transfected with pre-miR-control or pre-miR-503. (F) Luminescence quantification of MDA-MB-231 cells incubated with exosomes from HUVECs transfected with pre-miR-control or pre-miR-503. Additionally, see Fig. S3.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Figure 3: Endothelial miR-503 impairs tumor growth in vitro(A) Invasion level of MDA-MB-231 cells transfected with pre-miR-control or pre-miR-503 and with anti-miR-control or anti-miR-503. (B) Luminescence quantification of MDA-MB-231 cells transfected with pre-miR-control or pre-miR-503 and with anti-miR-control or anti-miR-503. (C) MiR-503 levels, measured by qRT-PCR in MDA-MB-231 cells incubated with 5 μg of HUVEC exosomes for 24 h. (D) Invasion level of MDA-MB-231 cells transfected with anti-miR-control or anti-miR-503 and cocultured with HUVECs transfected with pre-miR-control or pre-miR-503. (E) Luminescence quantification of MDA-MB-231 cells transfected with anti-miR-control or anti-miR-503 and cocultured with HUVECs transfected with pre-miR-control or pre-miR-503. (F) Luminescence quantification of MDA-MB-231 cells incubated with exosomes from HUVECs transfected with pre-miR-control or pre-miR-503. Additionally, see Fig. S3.
Mentions: To investigate the impact of miR-503 on tumor growth, we performed gain- and loss-of-function studies. MDA-MB-231 cells were transfected with either pre- or anti-miR-503, and the transfection efficiency was monitored using qRT-PCR (Fig. S3A). We used MDA-MB-231 constitutively expressing luciferase to quantify the proliferation in a coculture system by measuring the luminescence. Moreover, tumor cell invasion was assessed by quantifying the sprouting of tumor spheroids in a 3D collagen matrix. Increasing miR-503 levels via the transfection of miRNA mimics (pre-miRs) in MDA-MB-231 cells decreased both cell proliferation and invasion. Conversely, inhibition of miR-503 via the transfection of miR-503 antisense LNAs (anti-miRs) resulted in increased levels both of these processes compared with those of the control (Fig. 3A-B). Moreover, the effects of modulating miR-503 on tumor proliferation and invasion were confirmed by measuring BrdU incorporation and performing Boyden chamber assays, respectively (Fig. S3B-C).

Bottom Line: The modulation of miR-503 in breast cancer cells altered their proliferative and invasive capacities.We then identified two targets of miR-503, CCND2 and CCND3.Moreover, we measured increased plasmatic miR-503 in breast cancer patients after neoadjuvant chemotherapy, which could be partly due to increased miRNA secretion by endothelial cells.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Molecular Angiogenesis, GIGA-R, University of Liège, Belgium.

ABSTRACT
The interaction between tumor cells and their microenvironment is an essential aspect of tumor development. Therefore, understanding how this microenvironment communicates with tumor cells is crucial for the development of new anti-cancer therapies. MicroRNAs (miRNAs) are small non-coding RNAs that inhibit gene expression. They are secreted into the extracellular medium in vesicles called exosomes, which allow communication between cells via the transfer of their cargo. Consequently, we hypothesized that circulating endothelial miRNAs could be transferred to tumor cells and modify their phenotype. Using exogenous miRNA, we demonstrated that endothelial cells can transfer miRNA to tumor cells via exosomes. Using miRNA profiling, we identified miR-503, which exhibited downregulated levels in exosomes released from endothelial cells cultured under tumoral conditions. The modulation of miR-503 in breast cancer cells altered their proliferative and invasive capacities. We then identified two targets of miR-503, CCND2 and CCND3. Moreover, we measured increased plasmatic miR-503 in breast cancer patients after neoadjuvant chemotherapy, which could be partly due to increased miRNA secretion by endothelial cells. Taken together, our data are the first to reveal the involvement of the endothelium in the modulation of tumor development via the secretion of circulating miR-503 in response to chemotherapy treatment.

No MeSH data available.


Related in: MedlinePlus