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Estrogen adversely affects the prognosis of patients with lung adenocarcinoma.

Hsu LH, Liu KJ, Tsai MF, Wu CR, Feng AC, Chu NM, Kao SH - Cancer Sci. (2014)

Bottom Line: We found that the premenopausal patients had more advanced disease and a shorter survival among the never-smoking female patients with lung adenocarcinoma.We proposed a different pathway that estrogen upregulated the expression of osteopontin and then promoted cell migration through αvβ3 integrin binding and activated MEK-ERK signaling pathway, which is a common downstream pathway with epidermal growth factor receptor (EGFR) activation.An additive effect of ER antagonists and EGFR antagonists on the inhibition of cell migration was also noted.

View Article: PubMed Central - PubMed

Affiliation: Division of Pulmonary and Critical Care Medicine, Sun Yat-Sen Cancer Center, Taipei, Taiwan; Department of Medicine, National Yang-Ming University Medical School, Taipei, Taiwan.

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Estrogen induced osteopontin (OPN) expression and promoted cell migration through αvβ3 integrin binding and MEK/ERK signaling. (a) Gel electrophoretogram of OPN (spp1) gene expression of A549 cells in the presence of E2 and/ or tamoxifen. (b) The expression levels of OPN mRNA were quantified by quantitative real-time PCR. **P < 0.01 compared with the control; #P < 0.05 compared with the E2 group. (c) Western blotting of OPN in the culture media from the cells treated with E2 and/or tamoxifen was analyzed. Both the E2-induced OPN mRNA and protein expression were reduced in the presence of tamoxifen. **P < 0.01 compared with the control; ##P < 0.01 compared with the E2 group. (d) Wound healing assay of A549 or PE089 cells incubated for 24 h with OPN and/or U0126 (U). U0126 inhibited OPN-promoted cell migration. *P < 0.05 compared with the control; #P < 0.05 compared with the OPN group. (e) Western blotting of ERK and phosphorylated ERK of the cells treated with OPN and/or U0126. OPN-induced ERK phosphorylation was inhibited by U0126. (f) Wound healing assay of A549 or PE089 cells incubated with OPN, E2, EGF with/without anti-αvβ3 antibody (αvβ3 Ab) or siOPN for 24 h. αvβ3 Ab and siOPN attenuated the E2 or EGF-induced cell migration. *P < 0.05 compared with the control; †P < 0.05 compared with the OPN group; #P < 0.05 compared with the E2 group; ¶P < 0.05 compared with the EGF group.
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fig04: Estrogen induced osteopontin (OPN) expression and promoted cell migration through αvβ3 integrin binding and MEK/ERK signaling. (a) Gel electrophoretogram of OPN (spp1) gene expression of A549 cells in the presence of E2 and/ or tamoxifen. (b) The expression levels of OPN mRNA were quantified by quantitative real-time PCR. **P < 0.01 compared with the control; #P < 0.05 compared with the E2 group. (c) Western blotting of OPN in the culture media from the cells treated with E2 and/or tamoxifen was analyzed. Both the E2-induced OPN mRNA and protein expression were reduced in the presence of tamoxifen. **P < 0.01 compared with the control; ##P < 0.01 compared with the E2 group. (d) Wound healing assay of A549 or PE089 cells incubated for 24 h with OPN and/or U0126 (U). U0126 inhibited OPN-promoted cell migration. *P < 0.05 compared with the control; #P < 0.05 compared with the OPN group. (e) Western blotting of ERK and phosphorylated ERK of the cells treated with OPN and/or U0126. OPN-induced ERK phosphorylation was inhibited by U0126. (f) Wound healing assay of A549 or PE089 cells incubated with OPN, E2, EGF with/without anti-αvβ3 antibody (αvβ3 Ab) or siOPN for 24 h. αvβ3 Ab and siOPN attenuated the E2 or EGF-induced cell migration. *P < 0.05 compared with the control; †P < 0.05 compared with the OPN group; #P < 0.05 compared with the E2 group; ¶P < 0.05 compared with the EGF group.

Mentions: To test whether OPN was a downstream molecule of the estrogen signaling pathway and whether it contributed to cell migration, the expression levels of the OPN gene were quantified. A 2.2-fold increase in OPN mRNA level was detected in the E2-treated A549 cells (Fig.4a,b). The protein levels of OPN also increased in the medium cultured with E2 (Fig.4c). Both the E2-induced OPN expressions were reduced in the presence of tamoxifen (Fig.4a–c). OPN-induced cell migration and ERK phosphorylation were significantly inhibited by U0126 (MAP kinase/MEK inhibitor) (Fig.4d,e). The integrin receptor αvβ3 interacts with OPN via the arginine-glycine-aspartate (RGD) motif. The anti-αvβ3 Ab significantly inhibited OPN-induced cell migration and E2-stimulated or EGF-stimulated cell migration (Fig.4f). These results suggested that estrogen induced OPN expression and promoted cancer cell migration through αvβ3 integrin binding and MEK/ERK signaling, and implied that OPN-αvβ3 integrin-induced cell migration was involved in the activation of ER and EGFR signaling pathways.


Estrogen adversely affects the prognosis of patients with lung adenocarcinoma.

Hsu LH, Liu KJ, Tsai MF, Wu CR, Feng AC, Chu NM, Kao SH - Cancer Sci. (2014)

Estrogen induced osteopontin (OPN) expression and promoted cell migration through αvβ3 integrin binding and MEK/ERK signaling. (a) Gel electrophoretogram of OPN (spp1) gene expression of A549 cells in the presence of E2 and/ or tamoxifen. (b) The expression levels of OPN mRNA were quantified by quantitative real-time PCR. **P < 0.01 compared with the control; #P < 0.05 compared with the E2 group. (c) Western blotting of OPN in the culture media from the cells treated with E2 and/or tamoxifen was analyzed. Both the E2-induced OPN mRNA and protein expression were reduced in the presence of tamoxifen. **P < 0.01 compared with the control; ##P < 0.01 compared with the E2 group. (d) Wound healing assay of A549 or PE089 cells incubated for 24 h with OPN and/or U0126 (U). U0126 inhibited OPN-promoted cell migration. *P < 0.05 compared with the control; #P < 0.05 compared with the OPN group. (e) Western blotting of ERK and phosphorylated ERK of the cells treated with OPN and/or U0126. OPN-induced ERK phosphorylation was inhibited by U0126. (f) Wound healing assay of A549 or PE089 cells incubated with OPN, E2, EGF with/without anti-αvβ3 antibody (αvβ3 Ab) or siOPN for 24 h. αvβ3 Ab and siOPN attenuated the E2 or EGF-induced cell migration. *P < 0.05 compared with the control; †P < 0.05 compared with the OPN group; #P < 0.05 compared with the E2 group; ¶P < 0.05 compared with the EGF group.
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig04: Estrogen induced osteopontin (OPN) expression and promoted cell migration through αvβ3 integrin binding and MEK/ERK signaling. (a) Gel electrophoretogram of OPN (spp1) gene expression of A549 cells in the presence of E2 and/ or tamoxifen. (b) The expression levels of OPN mRNA were quantified by quantitative real-time PCR. **P < 0.01 compared with the control; #P < 0.05 compared with the E2 group. (c) Western blotting of OPN in the culture media from the cells treated with E2 and/or tamoxifen was analyzed. Both the E2-induced OPN mRNA and protein expression were reduced in the presence of tamoxifen. **P < 0.01 compared with the control; ##P < 0.01 compared with the E2 group. (d) Wound healing assay of A549 or PE089 cells incubated for 24 h with OPN and/or U0126 (U). U0126 inhibited OPN-promoted cell migration. *P < 0.05 compared with the control; #P < 0.05 compared with the OPN group. (e) Western blotting of ERK and phosphorylated ERK of the cells treated with OPN and/or U0126. OPN-induced ERK phosphorylation was inhibited by U0126. (f) Wound healing assay of A549 or PE089 cells incubated with OPN, E2, EGF with/without anti-αvβ3 antibody (αvβ3 Ab) or siOPN for 24 h. αvβ3 Ab and siOPN attenuated the E2 or EGF-induced cell migration. *P < 0.05 compared with the control; †P < 0.05 compared with the OPN group; #P < 0.05 compared with the E2 group; ¶P < 0.05 compared with the EGF group.
Mentions: To test whether OPN was a downstream molecule of the estrogen signaling pathway and whether it contributed to cell migration, the expression levels of the OPN gene were quantified. A 2.2-fold increase in OPN mRNA level was detected in the E2-treated A549 cells (Fig.4a,b). The protein levels of OPN also increased in the medium cultured with E2 (Fig.4c). Both the E2-induced OPN expressions were reduced in the presence of tamoxifen (Fig.4a–c). OPN-induced cell migration and ERK phosphorylation were significantly inhibited by U0126 (MAP kinase/MEK inhibitor) (Fig.4d,e). The integrin receptor αvβ3 interacts with OPN via the arginine-glycine-aspartate (RGD) motif. The anti-αvβ3 Ab significantly inhibited OPN-induced cell migration and E2-stimulated or EGF-stimulated cell migration (Fig.4f). These results suggested that estrogen induced OPN expression and promoted cancer cell migration through αvβ3 integrin binding and MEK/ERK signaling, and implied that OPN-αvβ3 integrin-induced cell migration was involved in the activation of ER and EGFR signaling pathways.

Bottom Line: We found that the premenopausal patients had more advanced disease and a shorter survival among the never-smoking female patients with lung adenocarcinoma.We proposed a different pathway that estrogen upregulated the expression of osteopontin and then promoted cell migration through αvβ3 integrin binding and activated MEK-ERK signaling pathway, which is a common downstream pathway with epidermal growth factor receptor (EGFR) activation.An additive effect of ER antagonists and EGFR antagonists on the inhibition of cell migration was also noted.

View Article: PubMed Central - PubMed

Affiliation: Division of Pulmonary and Critical Care Medicine, Sun Yat-Sen Cancer Center, Taipei, Taiwan; Department of Medicine, National Yang-Ming University Medical School, Taipei, Taiwan.

Show MeSH
Related in: MedlinePlus