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mPGES-1 in prostate cancer controls stemness and amplifies epidermal growth factor receptor-driven oncogenicity.

Finetti F, Terzuoli E, Giachetti A, Santi R, Villari D, Hanaka H, Radmark O, Ziche M, Donnini S - Endocr. Relat. Cancer (2015)

Bottom Line: There is evidence that an inflammatory microenvironment is associated with the development and progression of prostate cancer (PCa), although the determinants of intrinsic inflammation in PCa cells are not completely understood.They also show increased capacity to survive irrespective of anchorage condition, and overexpress EGFR compared to mPGES-1(KD) cells. mPGES-1 expression correlates with increased in vivo tumour growth and metastasis.Although EGFR inhibition reduces mPGES-1(SC) and mPGES-1(KD) cell xenograft tumour growth, we show that mPGES-1/PGE2 signalling sensitizes tumour cells to EGFR inhibitors.

View Article: PubMed Central - PubMed

Affiliation: Department of Life SciencesUniversity of Siena, Via Aldo Moro 2, 53100 Siena, ItalyDepartment of Surgery and Translational MedicineUniversity of Florence, Largo Brambilla 3, 50134 Firenze, ItalyDepartment of Clinical and Experimental MedicineUniversity of Florence, Viale Pieraccini 18, 50139 Firenze, ItalyDepartment of Medical Biochemistry and BiophysicsKarolinska Institutet, SE-171 77 Stockholm, SwedenIstituto Toscano Tumori (ITT)Firenze, Italy.

No MeSH data available.


Related in: MedlinePlus

mPGES-1 expression in cells mediates prostate tumor responsiveness to erlotinib. (A) Western blot analysis of the EGFR expression in DU145 mPGES-1SC and mPGES-1KD cells in basal condition or treated for 96 h with 1 μmol/l PGE2. Graph: quantification of EGFR expression. Data (three experiments) represent the ratio between EGFR and actin. **P<0.01 vs mPGES1SC and ##P<0.01 vs mPGES-1KD. (B) Immunofluorescence analysis of EGFR expression in mPGES-1SC and mPGES-1KD cells. (C and D) EGFR expression and ERK 1/2 phosphorylation in tumor specimens derived from DU145 mPGES-1SC and mPGES-1KD. (E and F) Western blot analysis of EGFR expression in DU145 and PC-3 mPGES-1SC treated with MF63 (10 μmol/l, 24–72 h), in DU145 mPGES-1SC treated with NS398 (10 μmol/l, 96 h) and in DU145 mPGES-1KD treated with PGE2 (1 μmol/l, 96 h). Representative gels of three experiments. (G) Western blot analysis of EGFR phosphorylation and vimentin expression in DU145 mPGES-1SC cells treated with erlotinib (10 μmol/l, 96 h). Data are representative of three experiments. (H) MTT assay of DU145 mPGES-1SC and mPGES-1KD cells treated with erlotinib (0.01–10 μmol/l, 96 h). Data (three experiments in triplicate) are reported as cell death (%). (I) Representative colonies of DU145 mPGES-1SC and mPGES-1KD cells. (J) Quantification of DU145 mPGES-1SC and mPGES-1KD colonies in the presence or absence of erlotinib 1 and 10 μmol/l. Data are reported as Δ of mPGES-1SC and mPGES-1KD cell colonies in the presence of erlotinib over the control (%) and are the means of three experiments in duplicate. *P<0.05 vs mPGES-1SC; **P<0.01 vs mPGES-1KD. (K) Tumor volume measured in athymic mice inoculated with DU145 mPGES-1SC or mPGES-1KD cells and treated with erlotinib (400 μg/mouse per three times per week, ip.). #P<0.05, **P<0.01 and ***P<0.001 vs DU145 mPGES-1SC.
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fig6: mPGES-1 expression in cells mediates prostate tumor responsiveness to erlotinib. (A) Western blot analysis of the EGFR expression in DU145 mPGES-1SC and mPGES-1KD cells in basal condition or treated for 96 h with 1 μmol/l PGE2. Graph: quantification of EGFR expression. Data (three experiments) represent the ratio between EGFR and actin. **P<0.01 vs mPGES1SC and ##P<0.01 vs mPGES-1KD. (B) Immunofluorescence analysis of EGFR expression in mPGES-1SC and mPGES-1KD cells. (C and D) EGFR expression and ERK 1/2 phosphorylation in tumor specimens derived from DU145 mPGES-1SC and mPGES-1KD. (E and F) Western blot analysis of EGFR expression in DU145 and PC-3 mPGES-1SC treated with MF63 (10 μmol/l, 24–72 h), in DU145 mPGES-1SC treated with NS398 (10 μmol/l, 96 h) and in DU145 mPGES-1KD treated with PGE2 (1 μmol/l, 96 h). Representative gels of three experiments. (G) Western blot analysis of EGFR phosphorylation and vimentin expression in DU145 mPGES-1SC cells treated with erlotinib (10 μmol/l, 96 h). Data are representative of three experiments. (H) MTT assay of DU145 mPGES-1SC and mPGES-1KD cells treated with erlotinib (0.01–10 μmol/l, 96 h). Data (three experiments in triplicate) are reported as cell death (%). (I) Representative colonies of DU145 mPGES-1SC and mPGES-1KD cells. (J) Quantification of DU145 mPGES-1SC and mPGES-1KD colonies in the presence or absence of erlotinib 1 and 10 μmol/l. Data are reported as Δ of mPGES-1SC and mPGES-1KD cell colonies in the presence of erlotinib over the control (%) and are the means of three experiments in duplicate. *P<0.05 vs mPGES-1SC; **P<0.01 vs mPGES-1KD. (K) Tumor volume measured in athymic mice inoculated with DU145 mPGES-1SC or mPGES-1KD cells and treated with erlotinib (400 μg/mouse per three times per week, ip.). #P<0.05, **P<0.01 and ***P<0.001 vs DU145 mPGES-1SC.

Mentions: It is well documented that PGE2 favours EGF/EGFR-induced oncogenicity by directly phosphorylating EGFR (Buchanan et al. 2003, Donnini et al. 2007). However, it is unknown whether mPGES-1/PGE2 signalling modulates EGFR expression levels.We measured EGFR expression in DU145 mPGES-1SC and mPGES-1KD cells, in tumours from mice inoculated with the respective cell lines and in DU145 and PC-3 treated with MF63. We found distinctly higher EGFR expression levels in the mPGES-1SC group than in the mPGES-1KD group (A, B and C and Supplementary Figure 2B). Consistently, p-ERK1/2, a known downstream effector of EGFR signalling, was higher in mPGES-1SC than mPGES-1KD tumours (Fig. 6D), suggesting that EGFR signalling plays a role in mPGES-1 enhancement of tumour growth.


mPGES-1 in prostate cancer controls stemness and amplifies epidermal growth factor receptor-driven oncogenicity.

Finetti F, Terzuoli E, Giachetti A, Santi R, Villari D, Hanaka H, Radmark O, Ziche M, Donnini S - Endocr. Relat. Cancer (2015)

mPGES-1 expression in cells mediates prostate tumor responsiveness to erlotinib. (A) Western blot analysis of the EGFR expression in DU145 mPGES-1SC and mPGES-1KD cells in basal condition or treated for 96 h with 1 μmol/l PGE2. Graph: quantification of EGFR expression. Data (three experiments) represent the ratio between EGFR and actin. **P<0.01 vs mPGES1SC and ##P<0.01 vs mPGES-1KD. (B) Immunofluorescence analysis of EGFR expression in mPGES-1SC and mPGES-1KD cells. (C and D) EGFR expression and ERK 1/2 phosphorylation in tumor specimens derived from DU145 mPGES-1SC and mPGES-1KD. (E and F) Western blot analysis of EGFR expression in DU145 and PC-3 mPGES-1SC treated with MF63 (10 μmol/l, 24–72 h), in DU145 mPGES-1SC treated with NS398 (10 μmol/l, 96 h) and in DU145 mPGES-1KD treated with PGE2 (1 μmol/l, 96 h). Representative gels of three experiments. (G) Western blot analysis of EGFR phosphorylation and vimentin expression in DU145 mPGES-1SC cells treated with erlotinib (10 μmol/l, 96 h). Data are representative of three experiments. (H) MTT assay of DU145 mPGES-1SC and mPGES-1KD cells treated with erlotinib (0.01–10 μmol/l, 96 h). Data (three experiments in triplicate) are reported as cell death (%). (I) Representative colonies of DU145 mPGES-1SC and mPGES-1KD cells. (J) Quantification of DU145 mPGES-1SC and mPGES-1KD colonies in the presence or absence of erlotinib 1 and 10 μmol/l. Data are reported as Δ of mPGES-1SC and mPGES-1KD cell colonies in the presence of erlotinib over the control (%) and are the means of three experiments in duplicate. *P<0.05 vs mPGES-1SC; **P<0.01 vs mPGES-1KD. (K) Tumor volume measured in athymic mice inoculated with DU145 mPGES-1SC or mPGES-1KD cells and treated with erlotinib (400 μg/mouse per three times per week, ip.). #P<0.05, **P<0.01 and ***P<0.001 vs DU145 mPGES-1SC.
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fig6: mPGES-1 expression in cells mediates prostate tumor responsiveness to erlotinib. (A) Western blot analysis of the EGFR expression in DU145 mPGES-1SC and mPGES-1KD cells in basal condition or treated for 96 h with 1 μmol/l PGE2. Graph: quantification of EGFR expression. Data (three experiments) represent the ratio between EGFR and actin. **P<0.01 vs mPGES1SC and ##P<0.01 vs mPGES-1KD. (B) Immunofluorescence analysis of EGFR expression in mPGES-1SC and mPGES-1KD cells. (C and D) EGFR expression and ERK 1/2 phosphorylation in tumor specimens derived from DU145 mPGES-1SC and mPGES-1KD. (E and F) Western blot analysis of EGFR expression in DU145 and PC-3 mPGES-1SC treated with MF63 (10 μmol/l, 24–72 h), in DU145 mPGES-1SC treated with NS398 (10 μmol/l, 96 h) and in DU145 mPGES-1KD treated with PGE2 (1 μmol/l, 96 h). Representative gels of three experiments. (G) Western blot analysis of EGFR phosphorylation and vimentin expression in DU145 mPGES-1SC cells treated with erlotinib (10 μmol/l, 96 h). Data are representative of three experiments. (H) MTT assay of DU145 mPGES-1SC and mPGES-1KD cells treated with erlotinib (0.01–10 μmol/l, 96 h). Data (three experiments in triplicate) are reported as cell death (%). (I) Representative colonies of DU145 mPGES-1SC and mPGES-1KD cells. (J) Quantification of DU145 mPGES-1SC and mPGES-1KD colonies in the presence or absence of erlotinib 1 and 10 μmol/l. Data are reported as Δ of mPGES-1SC and mPGES-1KD cell colonies in the presence of erlotinib over the control (%) and are the means of three experiments in duplicate. *P<0.05 vs mPGES-1SC; **P<0.01 vs mPGES-1KD. (K) Tumor volume measured in athymic mice inoculated with DU145 mPGES-1SC or mPGES-1KD cells and treated with erlotinib (400 μg/mouse per three times per week, ip.). #P<0.05, **P<0.01 and ***P<0.001 vs DU145 mPGES-1SC.
Mentions: It is well documented that PGE2 favours EGF/EGFR-induced oncogenicity by directly phosphorylating EGFR (Buchanan et al. 2003, Donnini et al. 2007). However, it is unknown whether mPGES-1/PGE2 signalling modulates EGFR expression levels.We measured EGFR expression in DU145 mPGES-1SC and mPGES-1KD cells, in tumours from mice inoculated with the respective cell lines and in DU145 and PC-3 treated with MF63. We found distinctly higher EGFR expression levels in the mPGES-1SC group than in the mPGES-1KD group (A, B and C and Supplementary Figure 2B). Consistently, p-ERK1/2, a known downstream effector of EGFR signalling, was higher in mPGES-1SC than mPGES-1KD tumours (Fig. 6D), suggesting that EGFR signalling plays a role in mPGES-1 enhancement of tumour growth.

Bottom Line: There is evidence that an inflammatory microenvironment is associated with the development and progression of prostate cancer (PCa), although the determinants of intrinsic inflammation in PCa cells are not completely understood.They also show increased capacity to survive irrespective of anchorage condition, and overexpress EGFR compared to mPGES-1(KD) cells. mPGES-1 expression correlates with increased in vivo tumour growth and metastasis.Although EGFR inhibition reduces mPGES-1(SC) and mPGES-1(KD) cell xenograft tumour growth, we show that mPGES-1/PGE2 signalling sensitizes tumour cells to EGFR inhibitors.

View Article: PubMed Central - PubMed

Affiliation: Department of Life SciencesUniversity of Siena, Via Aldo Moro 2, 53100 Siena, ItalyDepartment of Surgery and Translational MedicineUniversity of Florence, Largo Brambilla 3, 50134 Firenze, ItalyDepartment of Clinical and Experimental MedicineUniversity of Florence, Viale Pieraccini 18, 50139 Firenze, ItalyDepartment of Medical Biochemistry and BiophysicsKarolinska Institutet, SE-171 77 Stockholm, SwedenIstituto Toscano Tumori (ITT)Firenze, Italy.

No MeSH data available.


Related in: MedlinePlus