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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 do not affect the ability of platelets to bind to or to induce EMT-like changes in SK-OV-3 cells. a Platelet adhesion to SK-OV-3 cells in the presence and absence of aspirin or 2MeSAMP was quantified based on the fluorescence detection of CD42b positive platelets (n = 5). b Results are expressed as fold change in mRNA expression in SK-OV-3 cells in the presence of platelets (black bar) and platelets pre-treated with 2MeSAMP (white bar) relative to cells grown in the absence of platelets for 24 h (n = 3). c Results are expressed as fold change in mRNA expression in SK-OV-3 cells in the presence of platelets (black bar) and platelets pre-treated with aspirin (white bar) relative to cells grown in the absence of platelets for 24 h (n = 3). Values are normalised to B2M (beta 2 microglobulin). Data shown are mean value + SD. Student’s t-test was performed to determine significant differences between expression levels of the EMT markers in the SK-OV-3 cells cultured with platelets compared with those cultured with platelets pre-treated with antiplatelet agents. No significant differences were identified (significance was defined as p ≤ 0.05)
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Fig5: Antiplatelet agents do not affect the ability of platelets to bind to or to induce EMT-like changes in SK-OV-3 cells. a Platelet adhesion to SK-OV-3 cells in the presence and absence of aspirin or 2MeSAMP was quantified based on the fluorescence detection of CD42b positive platelets (n = 5). b Results are expressed as fold change in mRNA expression in SK-OV-3 cells in the presence of platelets (black bar) and platelets pre-treated with 2MeSAMP (white bar) relative to cells grown in the absence of platelets for 24 h (n = 3). c Results are expressed as fold change in mRNA expression in SK-OV-3 cells in the presence of platelets (black bar) and platelets pre-treated with aspirin (white bar) relative to cells grown in the absence of platelets for 24 h (n = 3). Values are normalised to B2M (beta 2 microglobulin). Data shown are mean value + SD. Student’s t-test was performed to determine significant differences between expression levels of the EMT markers in the SK-OV-3 cells cultured with platelets compared with those cultured with platelets pre-treated with antiplatelet agents. No significant differences were identified (significance was defined as p ≤ 0.05)

Mentions: We next sought to determine whether antiplatelet agents could inhibit platelets from cloaking and inducing EMT-like molecular changes in SK-OV-3 cells. The degree of platelet cloaking was measured based on the fluorescent detection of labelled platelets on the surface of cancer cells post incubation. The ability of platelets to adhere to SK-OV-3 cells was not diminished in the presence of either aspirin or 2MeSAMP (Fig. 5a). xFurthermore, analysis of the gene expression profile of the 11 selected EMT markers in SK-OV-3 cells incubated with drug treated and untreated platelets for 24 h were preformed. While pre-treatment of platelets with antiplatelet agents diminishes the platelet-induced invasion capacity of SK-OV-3 cells, it did not inhibit their ability to induce gene expression alterations in EMT-related genes. There were no significant differences in the expression values between SK-OV-3 cells exposed to platelets and those exposed to platelets pre-treated with either antiplatelet agent (Fig. 5b and c).Fig. 5


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 do not affect the ability of platelets to bind to or to induce EMT-like changes in SK-OV-3 cells. a Platelet adhesion to SK-OV-3 cells in the presence and absence of aspirin or 2MeSAMP was quantified based on the fluorescence detection of CD42b positive platelets (n = 5). b Results are expressed as fold change in mRNA expression in SK-OV-3 cells in the presence of platelets (black bar) and platelets pre-treated with 2MeSAMP (white bar) relative to cells grown in the absence of platelets for 24 h (n = 3). c Results are expressed as fold change in mRNA expression in SK-OV-3 cells in the presence of platelets (black bar) and platelets pre-treated with aspirin (white bar) relative to cells grown in the absence of platelets for 24 h (n = 3). Values are normalised to B2M (beta 2 microglobulin). Data shown are mean value + SD. Student’s t-test was performed to determine significant differences between expression levels of the EMT markers in the SK-OV-3 cells cultured with platelets compared with those cultured with platelets pre-treated with antiplatelet agents. No significant differences were identified (significance was defined as p ≤ 0.05)
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Related In: Results  -  Collection

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Fig5: Antiplatelet agents do not affect the ability of platelets to bind to or to induce EMT-like changes in SK-OV-3 cells. a Platelet adhesion to SK-OV-3 cells in the presence and absence of aspirin or 2MeSAMP was quantified based on the fluorescence detection of CD42b positive platelets (n = 5). b Results are expressed as fold change in mRNA expression in SK-OV-3 cells in the presence of platelets (black bar) and platelets pre-treated with 2MeSAMP (white bar) relative to cells grown in the absence of platelets for 24 h (n = 3). c Results are expressed as fold change in mRNA expression in SK-OV-3 cells in the presence of platelets (black bar) and platelets pre-treated with aspirin (white bar) relative to cells grown in the absence of platelets for 24 h (n = 3). Values are normalised to B2M (beta 2 microglobulin). Data shown are mean value + SD. Student’s t-test was performed to determine significant differences between expression levels of the EMT markers in the SK-OV-3 cells cultured with platelets compared with those cultured with platelets pre-treated with antiplatelet agents. No significant differences were identified (significance was defined as p ≤ 0.05)
Mentions: We next sought to determine whether antiplatelet agents could inhibit platelets from cloaking and inducing EMT-like molecular changes in SK-OV-3 cells. The degree of platelet cloaking was measured based on the fluorescent detection of labelled platelets on the surface of cancer cells post incubation. The ability of platelets to adhere to SK-OV-3 cells was not diminished in the presence of either aspirin or 2MeSAMP (Fig. 5a). xFurthermore, analysis of the gene expression profile of the 11 selected EMT markers in SK-OV-3 cells incubated with drug treated and untreated platelets for 24 h were preformed. While pre-treatment of platelets with antiplatelet agents diminishes the platelet-induced invasion capacity of SK-OV-3 cells, it did not inhibit their ability to induce gene expression alterations in EMT-related genes. There were no significant differences in the expression values between SK-OV-3 cells exposed to platelets and those exposed to platelets pre-treated with either antiplatelet agent (Fig. 5b and c).Fig. 5

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