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Aspirin and P2Y12 inhibition attenuate platelet-induced ovarian cancer cell invasion.

Cooke NM, Spillane CD, Sheils O, O'Leary J, Kenny D - BMC Cancer (2015)

Bottom Line: Platelet-cancer cell interactions play a key role in successful haematogenous metastasis.Both aspirin (p ≤ 0.05) and 2-methylthioadenosine 5'-monophosphate triethylammonium salt hydrate (P2Y12 inhibitor; p ≤ 0.01) significantly decreased their invasion capacity, and effectively reverted invasion to levels comparable to SK-OV-3 cells alone.While there is increasing evidence for the cancer-protective effect of aspirin, this study suggests P2Y12 inhibition may also play a role.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular and Cellular Therapeutics, Royal College of Surgeons in Ireland, 123 St Stephens Green, Dublin 2, Ireland. niamhcooke@rcsi.ie.

ABSTRACT

Background: Platelet-cancer cell interactions play a key role in successful haematogenous metastasis. Disseminated malignancy is the leading cause of death among ovarian cancer patients. It is unknown why different ovarian cancers have different metastatic phenotypes. To investigate if platelet-cancer cell interactions play a role, we characterized the response of ovarian cancer cell lines to platelets both functionally and at a molecular level.

Methods: Cell lines 59 M and SK-OV-3 were used as in vitro model systems of metastatic ovarian cancer. Platelet cloaking of each cell line was quantified by flow cytometry. Matrigel invasion chamber assays were used to assess the invasive capacity of the cell lines. The induction of an EMT was assessed by morphology analysis and by gene expression analysis of a panel of 11 EMT markers using TaqMan RT-PCR.

Results: SK-OV-3 cells adhered to and activated more platelets than 59 M cells (p = 0.0333). Platelets significantly promoted the ability of only SK-OV-3 cells to invade (p ≤ 0.0001). Morphology and transcritpome analysis indicated that platelets induce an epithelial-to-mesenchymal transition phenotype in both cells lines, with a more exaggerated response in SK-OV-3 cells. Next, we investigated if antiplatelet agents could abrogate the platelet-induced aggressive phenotype in SK-OV-3 cells. Both aspirin (p ≤ 0.05) and 2-methylthioadenosine 5'-monophosphate triethylammonium salt hydrate (P2Y12 inhibitor; p ≤ 0.01) significantly decreased their invasion capacity, and effectively reverted invasion to levels comparable to SK-OV-3 cells alone.

Conclusion: While there is increasing evidence for the cancer-protective effect of aspirin, this study suggests P2Y12 inhibition may also play a role. Understanding these complex interactions between platelets and cancer cells could ultimately allow the establishment of therapies tailored to inhibiting metastasis, thus significantly reducing cancer morbidity.

No MeSH data available.


Related in: MedlinePlus

Antiplatelet agents significantly reduce platelet-mediated SK-OV-3 cell invasion. Results are expressed as the percentage of SK-OV-3 cell invasion, in the absence and presence of (a) 2MeSAMP-treated platelets (plts) and (b) aspirin-treated platelets, through Matrigel® invasion chambers relative to control chambers after 16 and 24 h (n = 4). Data shown are mean value + SD. *** p ≤ 0.001, ** p ≤ 0.01, * p ≤ 0.05 was determined by Student’s t test
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Fig4: Antiplatelet agents significantly reduce platelet-mediated SK-OV-3 cell invasion. Results are expressed as the percentage of SK-OV-3 cell invasion, in the absence and presence of (a) 2MeSAMP-treated platelets (plts) and (b) aspirin-treated platelets, through Matrigel® invasion chambers relative to control chambers after 16 and 24 h (n = 4). Data shown are mean value + SD. *** p ≤ 0.001, ** p ≤ 0.01, * p ≤ 0.05 was determined by Student’s t test

Mentions: 2MeSAMP inhibits platelet function by increasing the levels of cyclic AMP and/or actively competing with ADP for the P2Y12 receptor [30]. Upon addition of platelets pre-treated with 50 μM 2MeSAMP, the invasion activity of SK-OV-3 cells significantly decreased 4.3- and 2.5-fold compared to those incubated with untreated platelets after 16 and 24 h respectively (p = 0.0065 and p = 0.0118, Fig. 4a). Prevention of ADP-induced platelet activation using 2MeSAMP effectively reverted platelet cloaked SK-OV-3 cell invasion to levels comparable to SK-OV-3 cells alone (11 % versus 13 %, and 14 % versus 17 %, after 16 h and 24 h, respectively). No significant changes in SK-OV-3 cell invasion activity were observed when cells were pre-treated with 2MeSAMP alone. Next we studied the effect of aspirin which irreversibly disrupts platelet function by inhibiting the COX-1 enzyme. Exposure of SK-OV-3 cells to platelets pre-treated with 20 μM aspirin resulted in a 2- and 1.6-fold significant reduction in their invasion activity compared to those incubated with untreated platelets after 16 and 24 h respectively (p = 0.012 and p = 0.0214, respectively, Fig. 4b). In contrast to P2Y12 inhibition, although COX-1 inhibition did significantly decrease the platelet-mediated stimulatory effect on the invasion activity of SK-OV-3 cells, it did not completely abolish the platelet effect. When compared with SK-OV-3 cells alone, those exposed to platelets pre-treated with aspirin had a significantly increased invasion activity after both 16 and 24 h (13 % versus 23 %; and 13 % versus 28 %, p = 0.028, respectively).Fig. 4


Aspirin and P2Y12 inhibition attenuate platelet-induced ovarian cancer cell invasion.

Cooke NM, Spillane CD, Sheils O, O'Leary J, Kenny D - BMC Cancer (2015)

Antiplatelet agents significantly reduce platelet-mediated SK-OV-3 cell invasion. Results are expressed as the percentage of SK-OV-3 cell invasion, in the absence and presence of (a) 2MeSAMP-treated platelets (plts) and (b) aspirin-treated platelets, through Matrigel® invasion chambers relative to control chambers after 16 and 24 h (n = 4). Data shown are mean value + SD. *** p ≤ 0.001, ** p ≤ 0.01, * p ≤ 0.05 was determined by Student’s t test
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4565001&req=5

Fig4: Antiplatelet agents significantly reduce platelet-mediated SK-OV-3 cell invasion. Results are expressed as the percentage of SK-OV-3 cell invasion, in the absence and presence of (a) 2MeSAMP-treated platelets (plts) and (b) aspirin-treated platelets, through Matrigel® invasion chambers relative to control chambers after 16 and 24 h (n = 4). Data shown are mean value + SD. *** p ≤ 0.001, ** p ≤ 0.01, * p ≤ 0.05 was determined by Student’s t test
Mentions: 2MeSAMP inhibits platelet function by increasing the levels of cyclic AMP and/or actively competing with ADP for the P2Y12 receptor [30]. Upon addition of platelets pre-treated with 50 μM 2MeSAMP, the invasion activity of SK-OV-3 cells significantly decreased 4.3- and 2.5-fold compared to those incubated with untreated platelets after 16 and 24 h respectively (p = 0.0065 and p = 0.0118, Fig. 4a). Prevention of ADP-induced platelet activation using 2MeSAMP effectively reverted platelet cloaked SK-OV-3 cell invasion to levels comparable to SK-OV-3 cells alone (11 % versus 13 %, and 14 % versus 17 %, after 16 h and 24 h, respectively). No significant changes in SK-OV-3 cell invasion activity were observed when cells were pre-treated with 2MeSAMP alone. Next we studied the effect of aspirin which irreversibly disrupts platelet function by inhibiting the COX-1 enzyme. Exposure of SK-OV-3 cells to platelets pre-treated with 20 μM aspirin resulted in a 2- and 1.6-fold significant reduction in their invasion activity compared to those incubated with untreated platelets after 16 and 24 h respectively (p = 0.012 and p = 0.0214, respectively, Fig. 4b). In contrast to P2Y12 inhibition, although COX-1 inhibition did significantly decrease the platelet-mediated stimulatory effect on the invasion activity of SK-OV-3 cells, it did not completely abolish the platelet effect. When compared with SK-OV-3 cells alone, those exposed to platelets pre-treated with aspirin had a significantly increased invasion activity after both 16 and 24 h (13 % versus 23 %; and 13 % versus 28 %, p = 0.028, respectively).Fig. 4

Bottom Line: Platelet-cancer cell interactions play a key role in successful haematogenous metastasis.Both aspirin (p ≤ 0.05) and 2-methylthioadenosine 5'-monophosphate triethylammonium salt hydrate (P2Y12 inhibitor; p ≤ 0.01) significantly decreased their invasion capacity, and effectively reverted invasion to levels comparable to SK-OV-3 cells alone.While there is increasing evidence for the cancer-protective effect of aspirin, this study suggests P2Y12 inhibition may also play a role.

View Article: PubMed Central - PubMed

Affiliation: Department of Molecular and Cellular Therapeutics, Royal College of Surgeons in Ireland, 123 St Stephens Green, Dublin 2, Ireland. niamhcooke@rcsi.ie.

ABSTRACT

Background: Platelet-cancer cell interactions play a key role in successful haematogenous metastasis. Disseminated malignancy is the leading cause of death among ovarian cancer patients. It is unknown why different ovarian cancers have different metastatic phenotypes. To investigate if platelet-cancer cell interactions play a role, we characterized the response of ovarian cancer cell lines to platelets both functionally and at a molecular level.

Methods: Cell lines 59 M and SK-OV-3 were used as in vitro model systems of metastatic ovarian cancer. Platelet cloaking of each cell line was quantified by flow cytometry. Matrigel invasion chamber assays were used to assess the invasive capacity of the cell lines. The induction of an EMT was assessed by morphology analysis and by gene expression analysis of a panel of 11 EMT markers using TaqMan RT-PCR.

Results: SK-OV-3 cells adhered to and activated more platelets than 59 M cells (p = 0.0333). Platelets significantly promoted the ability of only SK-OV-3 cells to invade (p ≤ 0.0001). Morphology and transcritpome analysis indicated that platelets induce an epithelial-to-mesenchymal transition phenotype in both cells lines, with a more exaggerated response in SK-OV-3 cells. Next, we investigated if antiplatelet agents could abrogate the platelet-induced aggressive phenotype in SK-OV-3 cells. Both aspirin (p ≤ 0.05) and 2-methylthioadenosine 5'-monophosphate triethylammonium salt hydrate (P2Y12 inhibitor; p ≤ 0.01) significantly decreased their invasion capacity, and effectively reverted invasion to levels comparable to SK-OV-3 cells alone.

Conclusion: While there is increasing evidence for the cancer-protective effect of aspirin, this study suggests P2Y12 inhibition may also play a role. Understanding these complex interactions between platelets and cancer cells could ultimately allow the establishment of therapies tailored to inhibiting metastasis, thus significantly reducing cancer morbidity.

No MeSH data available.


Related in: MedlinePlus