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PinX1 inhibits the invasion and metastasis of human breast cancer via suppressing NF-κB/MMP-9 signaling pathway.

Shi M, Cao M, Song J, Liu Q, Li H, Meng F, Pan Z, Bai J, Zheng J - Mol. Cancer (2015)

Bottom Line: PinX1 (PIN2/TRF1-interacting telomerase inhibitor 1) was suggested to be correlated with tumor progression.We found that low PinX1 expression was associated with lymph node metastasis (P = 0.002) and histology grade (P = 0.001) in patients, as well as with poorer overall and disease-specific survival (P = 0.010 and P = 0.003, respectively).Finally, our mice model confirmed that PinX1 suppressed breast cancer metastasis in vivo.

View Article: PubMed Central - PubMed

Affiliation: Jiangsu Center for the Collaboration and Innovation of Cancer Biotherapy, Cancer Institute, Xuzhou Medical College, 84 West Huaihai Road, Xuzhou, 221002, Jiangsu Province, China. meilin-1018@163.com.

ABSTRACT

Background: PinX1 (PIN2/TRF1-interacting telomerase inhibitor 1) was suggested to be correlated with tumor progression. This study was designed to evaluate the role of PinX1 in human breast cancer.

Methods: To evaluate the function of PinX1 in breast cancer, we used a tissue microarray (TMA) of 405 human breast cancer patients and immunohistochemistry to analyze the correlation between PinX1 expression and clinicopathologic variables and patient survival. We also detected the abilities of cell migration and invasion in breast cancer by performing cell migration and invasion assay, gelatin zymography and western blot analysis. Lastly, we set up the nude mice model by Tail vein assay to exam the functional role of PinX1 in breast cancer metastasis.

Results: We found that low PinX1 expression was associated with lymph node metastasis (P = 0.002) and histology grade (P = 0.001) in patients, as well as with poorer overall and disease-specific survival (P = 0.010 and P = 0.003, respectively). Moreover, we identified that PinX1 inhibited the migration and invasion of breast cancer by suppressing MMP-9 expression and activity via NF-κB-dependent transcription in vitro. Finally, our mice model confirmed that PinX1 suppressed breast cancer metastasis in vivo.

Conclusions: Our data revealed that low PinX1 expression was an independent negative prognostic factor for breast cancer patients. These findings suggested that PinX1 might be function as a tumor metastasis suppressor in the development and progression of breast cancer by regulating the NF-κB/MMP-9 signaling pathway, and might be a prognostic marker as well as a therapeutic target for breast cancer.

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PinX1 inhibits migration and invasion of human breast cancer cells. (A) Western blot analysis of the relative protein level of PinX1 in PinX1 knockdown (siPinX1) and control siRNA (siCtrl) groups for both MDA-MB-231 and BT-549 cell lines. (B) Western blot analysis of the relative protein level of PinX1 in PinX1 overexpression (PinX1OE) and control vector (Vector) groups for both MDA-MB-231 and BT-549 cell lines. (C) and (E) PinX1 knockdown significantly inhibited migration and invasion abilities of MDA-MB-231 and BT-549 cells. (D) and (F) PinX1 overexpression significantly inhibited migration and invasion abilities of MDA-MB-231 and BT-549 cells. All experiments were carried out in triplicate. Data are shown as means ± SD. ***, P < 0.001.
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Fig2: PinX1 inhibits migration and invasion of human breast cancer cells. (A) Western blot analysis of the relative protein level of PinX1 in PinX1 knockdown (siPinX1) and control siRNA (siCtrl) groups for both MDA-MB-231 and BT-549 cell lines. (B) Western blot analysis of the relative protein level of PinX1 in PinX1 overexpression (PinX1OE) and control vector (Vector) groups for both MDA-MB-231 and BT-549 cell lines. (C) and (E) PinX1 knockdown significantly inhibited migration and invasion abilities of MDA-MB-231 and BT-549 cells. (D) and (F) PinX1 overexpression significantly inhibited migration and invasion abilities of MDA-MB-231 and BT-549 cells. All experiments were carried out in triplicate. Data are shown as means ± SD. ***, P < 0.001.

Mentions: Because low PinX1 expression is associated with poor prognosis, supporting PinX1 may play important roles in one or more steps of tumor metastasis. We investigated the involvement of PinX1 in breast cancer cells migration and invasion. We transiently transfected MDA-MB-231 and BT-549 cells with control siRNA and PinX1 siRNA or pEGFP-C3 and pEGFP-C3-PinX1 plasmids, respectively. Forty-eight or twenty-four hours after transfection, PinX1 protein was significantly knockdown or overexpressed in cancer cells, respectively (Figure 2A & B). In cell migration assay, we found that the ability of cell migration was drastically increased after PinX1 knockdown in both MDA-MB-231 and BT-549 cell line (Figure 2C). In contrast, overexpression of PinX1 inhibited cell migration (Figure 2D). Meanwhile, the results of the cell invasion assay corresponded with the cell migration assay (Figure 2E & F). However, overexpression or knockdown of PinX1 had no detectable effect on the proliferation of breast cancer cells under normal culture conditions (data not shown).Figure 2


PinX1 inhibits the invasion and metastasis of human breast cancer via suppressing NF-κB/MMP-9 signaling pathway.

Shi M, Cao M, Song J, Liu Q, Li H, Meng F, Pan Z, Bai J, Zheng J - Mol. Cancer (2015)

PinX1 inhibits migration and invasion of human breast cancer cells. (A) Western blot analysis of the relative protein level of PinX1 in PinX1 knockdown (siPinX1) and control siRNA (siCtrl) groups for both MDA-MB-231 and BT-549 cell lines. (B) Western blot analysis of the relative protein level of PinX1 in PinX1 overexpression (PinX1OE) and control vector (Vector) groups for both MDA-MB-231 and BT-549 cell lines. (C) and (E) PinX1 knockdown significantly inhibited migration and invasion abilities of MDA-MB-231 and BT-549 cells. (D) and (F) PinX1 overexpression significantly inhibited migration and invasion abilities of MDA-MB-231 and BT-549 cells. All experiments were carried out in triplicate. Data are shown as means ± SD. ***, P < 0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4404090&req=5

Fig2: PinX1 inhibits migration and invasion of human breast cancer cells. (A) Western blot analysis of the relative protein level of PinX1 in PinX1 knockdown (siPinX1) and control siRNA (siCtrl) groups for both MDA-MB-231 and BT-549 cell lines. (B) Western blot analysis of the relative protein level of PinX1 in PinX1 overexpression (PinX1OE) and control vector (Vector) groups for both MDA-MB-231 and BT-549 cell lines. (C) and (E) PinX1 knockdown significantly inhibited migration and invasion abilities of MDA-MB-231 and BT-549 cells. (D) and (F) PinX1 overexpression significantly inhibited migration and invasion abilities of MDA-MB-231 and BT-549 cells. All experiments were carried out in triplicate. Data are shown as means ± SD. ***, P < 0.001.
Mentions: Because low PinX1 expression is associated with poor prognosis, supporting PinX1 may play important roles in one or more steps of tumor metastasis. We investigated the involvement of PinX1 in breast cancer cells migration and invasion. We transiently transfected MDA-MB-231 and BT-549 cells with control siRNA and PinX1 siRNA or pEGFP-C3 and pEGFP-C3-PinX1 plasmids, respectively. Forty-eight or twenty-four hours after transfection, PinX1 protein was significantly knockdown or overexpressed in cancer cells, respectively (Figure 2A & B). In cell migration assay, we found that the ability of cell migration was drastically increased after PinX1 knockdown in both MDA-MB-231 and BT-549 cell line (Figure 2C). In contrast, overexpression of PinX1 inhibited cell migration (Figure 2D). Meanwhile, the results of the cell invasion assay corresponded with the cell migration assay (Figure 2E & F). However, overexpression or knockdown of PinX1 had no detectable effect on the proliferation of breast cancer cells under normal culture conditions (data not shown).Figure 2

Bottom Line: PinX1 (PIN2/TRF1-interacting telomerase inhibitor 1) was suggested to be correlated with tumor progression.We found that low PinX1 expression was associated with lymph node metastasis (P = 0.002) and histology grade (P = 0.001) in patients, as well as with poorer overall and disease-specific survival (P = 0.010 and P = 0.003, respectively).Finally, our mice model confirmed that PinX1 suppressed breast cancer metastasis in vivo.

View Article: PubMed Central - PubMed

Affiliation: Jiangsu Center for the Collaboration and Innovation of Cancer Biotherapy, Cancer Institute, Xuzhou Medical College, 84 West Huaihai Road, Xuzhou, 221002, Jiangsu Province, China. meilin-1018@163.com.

ABSTRACT

Background: PinX1 (PIN2/TRF1-interacting telomerase inhibitor 1) was suggested to be correlated with tumor progression. This study was designed to evaluate the role of PinX1 in human breast cancer.

Methods: To evaluate the function of PinX1 in breast cancer, we used a tissue microarray (TMA) of 405 human breast cancer patients and immunohistochemistry to analyze the correlation between PinX1 expression and clinicopathologic variables and patient survival. We also detected the abilities of cell migration and invasion in breast cancer by performing cell migration and invasion assay, gelatin zymography and western blot analysis. Lastly, we set up the nude mice model by Tail vein assay to exam the functional role of PinX1 in breast cancer metastasis.

Results: We found that low PinX1 expression was associated with lymph node metastasis (P = 0.002) and histology grade (P = 0.001) in patients, as well as with poorer overall and disease-specific survival (P = 0.010 and P = 0.003, respectively). Moreover, we identified that PinX1 inhibited the migration and invasion of breast cancer by suppressing MMP-9 expression and activity via NF-κB-dependent transcription in vitro. Finally, our mice model confirmed that PinX1 suppressed breast cancer metastasis in vivo.

Conclusions: Our data revealed that low PinX1 expression was an independent negative prognostic factor for breast cancer patients. These findings suggested that PinX1 might be function as a tumor metastasis suppressor in the development and progression of breast cancer by regulating the NF-κB/MMP-9 signaling pathway, and might be a prognostic marker as well as a therapeutic target for breast cancer.

Show MeSH
Related in: MedlinePlus