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Aquaporin3 is required for FGF-2-induced migration of human breast cancers.

Cao XC, Zhang WR, Cao WF, Liu BW, Zhang F, Zhao HM, Meng R, Zhang L, Niu RF, Hao XS, Zhang B - PLoS ONE (2013)

Bottom Line: The FGFR kinase inhibitor PD173074, significantly inhibited FGF-2-induced AQP3 expression and cell migration.The PI3K inhibitor LY294002 and MEK1/2 inhibitor PD98059 inhibited, but not fully blocked, FGF-2-induced AQP3 expression and cell migration.Our findings also suggest the importance of FGFR-PI3K and FGFR-ERK signaling in FGF-2-induced AQP3 expression.

View Article: PubMed Central - PubMed

Affiliation: National Key Laboratory of Breast Cancer Prevention and Treatment, Cancer Institute and Hospital, Tianjin, China.

ABSTRACT

Purpose: The aquaporin (AQP) family consists of a number of small integral membrane proteins that transport water and glycerol. AQPs are critical for trans-epithelial fluid transport. Recent reports demonstrated that AQPs, particularly AQP1 and AQP5, are expressed in high grade tumor cells of a variety of tissue origins, and that AQPs are involved in cell migration and metastasis. Based on this background, we examined whether AQP3, another important member of the AQP family, could facilitate cell migration in human breast cancers.

Methods: Potential role of AQP3 was examined using two representative breast cancer cell lines (MDA-MB-231 and Bcap-37). Briefly, AQP3 expression was inhibited with a lentivirus construct that stably expressed shRNA against the AQP3 mRNA. AQP3 expression inhibition was verified with Western blot. Cell migration was examined using a wound scratch assay in the presence of fibroblast growth factor-2 (FGF-2). In additional experiments, AQP3 was inhibited by CuSO4. Fibroblast growth factor receptor (FGFR) kinase inhibitor PD173074, PI3K inhibitor LY294002, and MEK1/2 inhibitor PD98059 were used to dissect the molecular mechanism of FGF-2 induced AQP3 expression.

Results: FGF-2 treatment increased AQP3 expression and induced cell migration in a dose dependent manner. Silencing AQP3 expression by a lentiviral shRNA inhibited FGF-2 induced cell migration. CuSO4, a water transport inhibitor selective for AQP3, also suppressed FGF-2-induced cell migration. The FGFR kinase inhibitor PD173074, significantly inhibited FGF-2-induced AQP3 expression and cell migration. The PI3K inhibitor LY294002 and MEK1/2 inhibitor PD98059 inhibited, but not fully blocked, FGF-2-induced AQP3 expression and cell migration.

Conclusions: AQP3 is required for FGF-2-induced cell migration in cultured human breast cancer cells. Our findings also suggest the importance of FGFR-PI3K and FGFR-ERK signaling in FGF-2-induced AQP3 expression. In summary, our findings suggest a novel function of AQP3 in cell migration and metastasis of breast cancers.

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PD173074 significantly inhibits FGF-2-induced AQP3 expression.MDA-MB-231 and Bcap-37 cells were treated with FGF-2 (10 ng/ml) with or without FGFR kinase inhibitor PD173074 (1 µM) for 24 h. AQP3 expression was analyzed by Western Blot and normalized to β-actin. The results represent means ± SEM for triplicate experiments. #P<0.05 versus untreated control (UNTR). *P<0.05 versus FGF-2 alone.
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pone-0056735-g006: PD173074 significantly inhibits FGF-2-induced AQP3 expression.MDA-MB-231 and Bcap-37 cells were treated with FGF-2 (10 ng/ml) with or without FGFR kinase inhibitor PD173074 (1 µM) for 24 h. AQP3 expression was analyzed by Western Blot and normalized to β-actin. The results represent means ± SEM for triplicate experiments. #P<0.05 versus untreated control (UNTR). *P<0.05 versus FGF-2 alone.

Mentions: Additional experiments were performed to study the signal pathway of FGF-2- induced AQP3 expression. To better comprehend the role of FGFR in FGF-2-induced AQP3 expression, PD173074, a FGFR kinase inhibitor, was used in the experiment. PD173074 significantly inhibited FGF-2-induced AQP3 expression (Figs. 6A and 6C). PD173074 decreased AQP3 expression by 74% and 90% in MDA-MB-231 and Bcap-37 cells, respectively (Figs. 6B and 6D). FGF-2-induced cell migration was also inhibited by PD173074 in both cell lines (Figs. 7A and 7C). The average gap was increased from 0.40-to 0.64-fold and from 0.28-to 0.57-fold in MDA-MB-231 and Bcap-37 cells, respectively (Figs. 7B and 7D).


Aquaporin3 is required for FGF-2-induced migration of human breast cancers.

Cao XC, Zhang WR, Cao WF, Liu BW, Zhang F, Zhao HM, Meng R, Zhang L, Niu RF, Hao XS, Zhang B - PLoS ONE (2013)

PD173074 significantly inhibits FGF-2-induced AQP3 expression.MDA-MB-231 and Bcap-37 cells were treated with FGF-2 (10 ng/ml) with or without FGFR kinase inhibitor PD173074 (1 µM) for 24 h. AQP3 expression was analyzed by Western Blot and normalized to β-actin. The results represent means ± SEM for triplicate experiments. #P<0.05 versus untreated control (UNTR). *P<0.05 versus FGF-2 alone.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3585269&req=5

pone-0056735-g006: PD173074 significantly inhibits FGF-2-induced AQP3 expression.MDA-MB-231 and Bcap-37 cells were treated with FGF-2 (10 ng/ml) with or without FGFR kinase inhibitor PD173074 (1 µM) for 24 h. AQP3 expression was analyzed by Western Blot and normalized to β-actin. The results represent means ± SEM for triplicate experiments. #P<0.05 versus untreated control (UNTR). *P<0.05 versus FGF-2 alone.
Mentions: Additional experiments were performed to study the signal pathway of FGF-2- induced AQP3 expression. To better comprehend the role of FGFR in FGF-2-induced AQP3 expression, PD173074, a FGFR kinase inhibitor, was used in the experiment. PD173074 significantly inhibited FGF-2-induced AQP3 expression (Figs. 6A and 6C). PD173074 decreased AQP3 expression by 74% and 90% in MDA-MB-231 and Bcap-37 cells, respectively (Figs. 6B and 6D). FGF-2-induced cell migration was also inhibited by PD173074 in both cell lines (Figs. 7A and 7C). The average gap was increased from 0.40-to 0.64-fold and from 0.28-to 0.57-fold in MDA-MB-231 and Bcap-37 cells, respectively (Figs. 7B and 7D).

Bottom Line: The FGFR kinase inhibitor PD173074, significantly inhibited FGF-2-induced AQP3 expression and cell migration.The PI3K inhibitor LY294002 and MEK1/2 inhibitor PD98059 inhibited, but not fully blocked, FGF-2-induced AQP3 expression and cell migration.Our findings also suggest the importance of FGFR-PI3K and FGFR-ERK signaling in FGF-2-induced AQP3 expression.

View Article: PubMed Central - PubMed

Affiliation: National Key Laboratory of Breast Cancer Prevention and Treatment, Cancer Institute and Hospital, Tianjin, China.

ABSTRACT

Purpose: The aquaporin (AQP) family consists of a number of small integral membrane proteins that transport water and glycerol. AQPs are critical for trans-epithelial fluid transport. Recent reports demonstrated that AQPs, particularly AQP1 and AQP5, are expressed in high grade tumor cells of a variety of tissue origins, and that AQPs are involved in cell migration and metastasis. Based on this background, we examined whether AQP3, another important member of the AQP family, could facilitate cell migration in human breast cancers.

Methods: Potential role of AQP3 was examined using two representative breast cancer cell lines (MDA-MB-231 and Bcap-37). Briefly, AQP3 expression was inhibited with a lentivirus construct that stably expressed shRNA against the AQP3 mRNA. AQP3 expression inhibition was verified with Western blot. Cell migration was examined using a wound scratch assay in the presence of fibroblast growth factor-2 (FGF-2). In additional experiments, AQP3 was inhibited by CuSO4. Fibroblast growth factor receptor (FGFR) kinase inhibitor PD173074, PI3K inhibitor LY294002, and MEK1/2 inhibitor PD98059 were used to dissect the molecular mechanism of FGF-2 induced AQP3 expression.

Results: FGF-2 treatment increased AQP3 expression and induced cell migration in a dose dependent manner. Silencing AQP3 expression by a lentiviral shRNA inhibited FGF-2 induced cell migration. CuSO4, a water transport inhibitor selective for AQP3, also suppressed FGF-2-induced cell migration. The FGFR kinase inhibitor PD173074, significantly inhibited FGF-2-induced AQP3 expression and cell migration. The PI3K inhibitor LY294002 and MEK1/2 inhibitor PD98059 inhibited, but not fully blocked, FGF-2-induced AQP3 expression and cell migration.

Conclusions: AQP3 is required for FGF-2-induced cell migration in cultured human breast cancer cells. Our findings also suggest the importance of FGFR-PI3K and FGFR-ERK signaling in FGF-2-induced AQP3 expression. In summary, our findings suggest a novel function of AQP3 in cell migration and metastasis of breast cancers.

Show MeSH
Related in: MedlinePlus