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CCR9 interactions support ovarian cancer cell survival and resistance to cisplatin-induced apoptosis in a PI3K-dependent and FAK-independent fashion.

Johnson EL, Singh R, Johnson-Holiday CM, Grizzle WE, Partridge EE, Lillard JW, Singh S - J Ovarian Res (2010)

Bottom Line: Our results show interactions between CCR9 and CCL25 increased anti-apoptotic signaling cascades in OvCa cells, which rescued cells from cisplatin-induced cell death.Specifically, CCL25-CCR9 interactions mediated Akt, activation as well as GSK-3beta and FKHR phosphorylation in a PI3K-dependent and FAK-independent fashion.Our results suggest the CCR9-CCL25 axis plays an important role in reducing cisplatin-induced apoptosis of OvCa cells.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Microbiology, Biochemistry and Immunology, Morehouse School of Medicine, 720 Westview Drive SW, Atlanta, GA 30310, USA. shsingh@msm.edu.

ABSTRACT

Background: Cisplatin is more often used to treat ovarian cancer (OvCa), which provides modest survival advantage primarily due to chemo-resistance and up regulated anti-apoptotic machineries in OvCa cells. Therefore, targeting the mechanisms responsible for cisplatin resistance in OvCa cell may improve therapeutic outcomes. We have shown that ovarian cancer cells express CC chemokine receptor-9 (CCR9). Others have also shown that CCL25, the only natural ligand for CCR9, up regulates anti-apoptotic proteins in immature T lymphocytes. Hence, it is plausible that CCR9-mediated cell signals might be involved in OvCa cell survival and inhibition of cisplatin-induced apoptosis. In this study, we investigated the potential role and molecular mechanisms of CCR9-mediated inhibition of cisplatin-induced apoptosis in OvCa cells.

Methods: Cell proliferation, vibrant apoptosis, and TUNEL assays were performed with or without cisplatin treatment in presence or absence of CCL25 to determine the role of the CCR9-CCL25 axis in cisplatin resistance. In situ Fast Activated cell-based ELISA (FACE) assays were performed to determine anti-apoptotic signaling molecules responsible for CCL25-CCR9 mediated survival.

Results: Our results show interactions between CCR9 and CCL25 increased anti-apoptotic signaling cascades in OvCa cells, which rescued cells from cisplatin-induced cell death. Specifically, CCL25-CCR9 interactions mediated Akt, activation as well as GSK-3beta and FKHR phosphorylation in a PI3K-dependent and FAK-independent fashion.

Conclusions: Our results suggest the CCR9-CCL25 axis plays an important role in reducing cisplatin-induced apoptosis of OvCa cells.

No MeSH data available.


Related in: MedlinePlus

CCL25 inhibits cisplatin-induced cell death. OVCAR-3 and SKOV-3 cells were cultured with 0 (circles) or 100 ng/ml of CCL25 plus isotype control (squares) or anti-CCR9 (triangles) antibodies for 24 hours, along with increasing concentrations of cisplatin. Cell proliferation was determined by BrdU incorporation and assays were repeated 3 times and performed in triplicate. Asterisk(s) (*) indicate statistical significant differences (p < 0.01) between CCL25-treated and untreated OvCa cells.
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Figure 1: CCL25 inhibits cisplatin-induced cell death. OVCAR-3 and SKOV-3 cells were cultured with 0 (circles) or 100 ng/ml of CCL25 plus isotype control (squares) or anti-CCR9 (triangles) antibodies for 24 hours, along with increasing concentrations of cisplatin. Cell proliferation was determined by BrdU incorporation and assays were repeated 3 times and performed in triplicate. Asterisk(s) (*) indicate statistical significant differences (p < 0.01) between CCL25-treated and untreated OvCa cells.

Mentions: SKOV-3 cells incorporated BrdU at a higher rate than OVCAR-3 cells (0.375 versus 0.250 OD405 nm, respectively), which suggested SKOV-3 cells proliferated at a higher rate compared to OVCAR-3 cells (Figure 1). In the absence of cisplatin, CCL25 significantly enhanced BrdU incorporation (i.e., growth) of OVCAR-3 and SKOV-3 cell lines by ~ 1.5-fold in comparison to untreated cells. However, when these cells were treated with increasing concentrations of cisplatin, CCL25 protected human OvCa cells from cisplatin-mediated growth inhibition. CCL25 optimally protected against 5 μg/ml or less cisplatin with 3.5 and 2.2-fold increases in OVCAR-3 and SKOV-3 cell BrdU incorporation respectively, in comparison to the untreated cells or CCL25 plus anti-CCR9 antibody treated cultures. In general, CCL25 treatment abrogated the growth inhibition of OVCAR-3 and SKOV-3 cell lines caused by cisplatin in a CCR9-dependent fashion.


CCR9 interactions support ovarian cancer cell survival and resistance to cisplatin-induced apoptosis in a PI3K-dependent and FAK-independent fashion.

Johnson EL, Singh R, Johnson-Holiday CM, Grizzle WE, Partridge EE, Lillard JW, Singh S - J Ovarian Res (2010)

CCL25 inhibits cisplatin-induced cell death. OVCAR-3 and SKOV-3 cells were cultured with 0 (circles) or 100 ng/ml of CCL25 plus isotype control (squares) or anti-CCR9 (triangles) antibodies for 24 hours, along with increasing concentrations of cisplatin. Cell proliferation was determined by BrdU incorporation and assays were repeated 3 times and performed in triplicate. Asterisk(s) (*) indicate statistical significant differences (p < 0.01) between CCL25-treated and untreated OvCa cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2914045&req=5

Figure 1: CCL25 inhibits cisplatin-induced cell death. OVCAR-3 and SKOV-3 cells were cultured with 0 (circles) or 100 ng/ml of CCL25 plus isotype control (squares) or anti-CCR9 (triangles) antibodies for 24 hours, along with increasing concentrations of cisplatin. Cell proliferation was determined by BrdU incorporation and assays were repeated 3 times and performed in triplicate. Asterisk(s) (*) indicate statistical significant differences (p < 0.01) between CCL25-treated and untreated OvCa cells.
Mentions: SKOV-3 cells incorporated BrdU at a higher rate than OVCAR-3 cells (0.375 versus 0.250 OD405 nm, respectively), which suggested SKOV-3 cells proliferated at a higher rate compared to OVCAR-3 cells (Figure 1). In the absence of cisplatin, CCL25 significantly enhanced BrdU incorporation (i.e., growth) of OVCAR-3 and SKOV-3 cell lines by ~ 1.5-fold in comparison to untreated cells. However, when these cells were treated with increasing concentrations of cisplatin, CCL25 protected human OvCa cells from cisplatin-mediated growth inhibition. CCL25 optimally protected against 5 μg/ml or less cisplatin with 3.5 and 2.2-fold increases in OVCAR-3 and SKOV-3 cell BrdU incorporation respectively, in comparison to the untreated cells or CCL25 plus anti-CCR9 antibody treated cultures. In general, CCL25 treatment abrogated the growth inhibition of OVCAR-3 and SKOV-3 cell lines caused by cisplatin in a CCR9-dependent fashion.

Bottom Line: Our results show interactions between CCR9 and CCL25 increased anti-apoptotic signaling cascades in OvCa cells, which rescued cells from cisplatin-induced cell death.Specifically, CCL25-CCR9 interactions mediated Akt, activation as well as GSK-3beta and FKHR phosphorylation in a PI3K-dependent and FAK-independent fashion.Our results suggest the CCR9-CCL25 axis plays an important role in reducing cisplatin-induced apoptosis of OvCa cells.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Microbiology, Biochemistry and Immunology, Morehouse School of Medicine, 720 Westview Drive SW, Atlanta, GA 30310, USA. shsingh@msm.edu.

ABSTRACT

Background: Cisplatin is more often used to treat ovarian cancer (OvCa), which provides modest survival advantage primarily due to chemo-resistance and up regulated anti-apoptotic machineries in OvCa cells. Therefore, targeting the mechanisms responsible for cisplatin resistance in OvCa cell may improve therapeutic outcomes. We have shown that ovarian cancer cells express CC chemokine receptor-9 (CCR9). Others have also shown that CCL25, the only natural ligand for CCR9, up regulates anti-apoptotic proteins in immature T lymphocytes. Hence, it is plausible that CCR9-mediated cell signals might be involved in OvCa cell survival and inhibition of cisplatin-induced apoptosis. In this study, we investigated the potential role and molecular mechanisms of CCR9-mediated inhibition of cisplatin-induced apoptosis in OvCa cells.

Methods: Cell proliferation, vibrant apoptosis, and TUNEL assays were performed with or without cisplatin treatment in presence or absence of CCL25 to determine the role of the CCR9-CCL25 axis in cisplatin resistance. In situ Fast Activated cell-based ELISA (FACE) assays were performed to determine anti-apoptotic signaling molecules responsible for CCL25-CCR9 mediated survival.

Results: Our results show interactions between CCR9 and CCL25 increased anti-apoptotic signaling cascades in OvCa cells, which rescued cells from cisplatin-induced cell death. Specifically, CCL25-CCR9 interactions mediated Akt, activation as well as GSK-3beta and FKHR phosphorylation in a PI3K-dependent and FAK-independent fashion.

Conclusions: Our results suggest the CCR9-CCL25 axis plays an important role in reducing cisplatin-induced apoptosis of OvCa cells.

No MeSH data available.


Related in: MedlinePlus