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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 breast cancer cells metastasis in vivo. (A) Left panel, Western blotting of PinX1 in there groups of Ctrl-MDA-MB-231 cell lines, PinX1OE-MDA-MB-231 cell lines and PinX1KD-MDA-MB-231 cell lines, which was selected with puromycin for 2 weeks after lentivirus infection. Right panel, without puromycin selection for 2 months, PinX1 expression levels were not changed in MDA-MB-231 stable cell lines. (B) H&E staining sections of lungs 2 months after injection of PinX1KD-MDA-MB-231 cell lines in BALB/c nude mouse through tail vein. Left panel, magnification × 100; right panel, magnification × 400. (C) Representative images of 10% buffered formalin fixed lungs with metastatic nodules 2 months after injection of Ctrl, PinX1OE and PinX1KD MDA-MB-231 stable cell lines. Arrows indicate metastatic nodules. (D) The number of lung metastatic nodules was counted under a dissecting microscope. Compared with the PinX1OE group, a statistically dramatic increase in the number of the lung metastases was seen in PinX1KD group, and these two groups also had significant diversity compared with Ctrl group respectively. Data are displayed with means ± SD from 10 mice in each group. ***, P < 0.001.
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Fig5: PinX1 inhibits breast cancer cells metastasis in vivo. (A) Left panel, Western blotting of PinX1 in there groups of Ctrl-MDA-MB-231 cell lines, PinX1OE-MDA-MB-231 cell lines and PinX1KD-MDA-MB-231 cell lines, which was selected with puromycin for 2 weeks after lentivirus infection. Right panel, without puromycin selection for 2 months, PinX1 expression levels were not changed in MDA-MB-231 stable cell lines. (B) H&E staining sections of lungs 2 months after injection of PinX1KD-MDA-MB-231 cell lines in BALB/c nude mouse through tail vein. Left panel, magnification × 100; right panel, magnification × 400. (C) Representative images of 10% buffered formalin fixed lungs with metastatic nodules 2 months after injection of Ctrl, PinX1OE and PinX1KD MDA-MB-231 stable cell lines. Arrows indicate metastatic nodules. (D) The number of lung metastatic nodules was counted under a dissecting microscope. Compared with the PinX1OE group, a statistically dramatic increase in the number of the lung metastases was seen in PinX1KD group, and these two groups also had significant diversity compared with Ctrl group respectively. Data are displayed with means ± SD from 10 mice in each group. ***, P < 0.001.

Mentions: Lastly, we examined whether PinX1 suppressed breast cancer metastasis in vivo. PinX1OE-MDA-MB-231, PinX1KD-MDA-MB-231 and Ctrl-MDA-MB-231 cell lines were established previously. After 3 weeks selection following with lentivirus infection, the PinX1 protein levels of these cell lines were confirmed by western blot. Then continuing to incubate them without adding puromycin for 2 months, we determined that the PinX1 protein expression levels of three stable cell lines had not been changed (Figure 5A).Figure 5


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 breast cancer cells metastasis in vivo. (A) Left panel, Western blotting of PinX1 in there groups of Ctrl-MDA-MB-231 cell lines, PinX1OE-MDA-MB-231 cell lines and PinX1KD-MDA-MB-231 cell lines, which was selected with puromycin for 2 weeks after lentivirus infection. Right panel, without puromycin selection for 2 months, PinX1 expression levels were not changed in MDA-MB-231 stable cell lines. (B) H&E staining sections of lungs 2 months after injection of PinX1KD-MDA-MB-231 cell lines in BALB/c nude mouse through tail vein. Left panel, magnification × 100; right panel, magnification × 400. (C) Representative images of 10% buffered formalin fixed lungs with metastatic nodules 2 months after injection of Ctrl, PinX1OE and PinX1KD MDA-MB-231 stable cell lines. Arrows indicate metastatic nodules. (D) The number of lung metastatic nodules was counted under a dissecting microscope. Compared with the PinX1OE group, a statistically dramatic increase in the number of the lung metastases was seen in PinX1KD group, and these two groups also had significant diversity compared with Ctrl group respectively. Data are displayed with means ± SD from 10 mice in each group. ***, P < 0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4404090&req=5

Fig5: PinX1 inhibits breast cancer cells metastasis in vivo. (A) Left panel, Western blotting of PinX1 in there groups of Ctrl-MDA-MB-231 cell lines, PinX1OE-MDA-MB-231 cell lines and PinX1KD-MDA-MB-231 cell lines, which was selected with puromycin for 2 weeks after lentivirus infection. Right panel, without puromycin selection for 2 months, PinX1 expression levels were not changed in MDA-MB-231 stable cell lines. (B) H&E staining sections of lungs 2 months after injection of PinX1KD-MDA-MB-231 cell lines in BALB/c nude mouse through tail vein. Left panel, magnification × 100; right panel, magnification × 400. (C) Representative images of 10% buffered formalin fixed lungs with metastatic nodules 2 months after injection of Ctrl, PinX1OE and PinX1KD MDA-MB-231 stable cell lines. Arrows indicate metastatic nodules. (D) The number of lung metastatic nodules was counted under a dissecting microscope. Compared with the PinX1OE group, a statistically dramatic increase in the number of the lung metastases was seen in PinX1KD group, and these two groups also had significant diversity compared with Ctrl group respectively. Data are displayed with means ± SD from 10 mice in each group. ***, P < 0.001.
Mentions: Lastly, we examined whether PinX1 suppressed breast cancer metastasis in vivo. PinX1OE-MDA-MB-231, PinX1KD-MDA-MB-231 and Ctrl-MDA-MB-231 cell lines were established previously. After 3 weeks selection following with lentivirus infection, the PinX1 protein levels of these cell lines were confirmed by western blot. Then continuing to incubate them without adding puromycin for 2 months, we determined that the PinX1 protein expression levels of three stable cell lines had not been changed (Figure 5A).Figure 5

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