Limits...
The Hippo/YAP pathway interacts with EGFR signaling and HPV oncoproteins to regulate cervical cancer progression.

He C, Mao D, Hua G, Lv X, Chen X, Angeletti PC, Dong J, Remmenga SW, Rodabaugh KJ, Zhou J, Lambert PF, Yang P, Davis JS, Wang C - EMBO Mol Med (2015)

Bottom Line: The Hippo signaling pathway controls organ size and tumorigenesis through a kinase cascade that inactivates Yes-associated protein (YAP).Our results suggest that YAP expression is associated with a poor prognosis for cervical cancer.Our study indicates that combined targeting of the Hippo and the ERBB signaling pathways represents a novel therapeutic strategy for prevention and treatment of cervical cancer.

View Article: PubMed Central - PubMed

Affiliation: Olson Center for Women's Health, Department of Obstetrics & Gynecology, University of Nebraska Medical Center, Omaha, NE, USA College of Animal Science and Technology, Huazhong Agricultural University, Wuhan, China.

Show MeSH

Related in: MedlinePlus

Function and expression of AREG in cervical cancer cellsWestern blot analysis showing that AREG treatment induced dephosphorylation of LATS1, MOB1, and YAP. ME180 cells were starved for 6 h after reaching cell confluence, then cells were treated with AREG (50 ng/ml) for 0, 30, 60 min.AREG (50 ng/ml) treatment induced appearance of elongated cells in ME180 cells (top and middle panels) and significantly stimulated ME180 cell proliferation (lower panel). Each bar represents mean ± SEM (n = 5). Bars with different letters are significantly different from each other (P = 0.003). Scale bar: 100 μm.Wound-healing assay showing that AREG stimulates migration of ME180 cervical cancer cells within 10 h in serum-free medium. Scale bar: 200 μm.Real-time PCR showing that treatment of ME180 cells with AREG for 24 h significantly increased AREG mRNA expression. Each bar represents mean ± SEM (n = 9). Bars with different letters are significantly different from each other (P < 0.0001).Real-time PCR showing that knockdown of YAP in ME180 cells with YAP siRNA (siYAP) significantly suppressed AREG-induced AREG mRNA expression. siCTRL (non-target siRNA) was used as a siRNA control. Each bar represents mean ± SEM (n = 4). Bars with different letters are significantly different from each other (siCTRL vs. siCTRL+AREG, P < 0.0001; siYAP+CTRL vs. siYAP+AREG, P < 0.0001; siCTRL+AREG vs. siYAP+AREG, P = 0.0037).Data information: Quantitative data in (B) and (D) were analyzed for significance using unpaired t-test in GraphPad Prism 5 with Welch’s correction. Data in (E) were analyzed for significance using one-way ANOVA in GraphPad Prism 5 with Tukey’s post hoc tests. Source data are available online for this figure.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4644376&req=5

fig07: Function and expression of AREG in cervical cancer cellsWestern blot analysis showing that AREG treatment induced dephosphorylation of LATS1, MOB1, and YAP. ME180 cells were starved for 6 h after reaching cell confluence, then cells were treated with AREG (50 ng/ml) for 0, 30, 60 min.AREG (50 ng/ml) treatment induced appearance of elongated cells in ME180 cells (top and middle panels) and significantly stimulated ME180 cell proliferation (lower panel). Each bar represents mean ± SEM (n = 5). Bars with different letters are significantly different from each other (P = 0.003). Scale bar: 100 μm.Wound-healing assay showing that AREG stimulates migration of ME180 cervical cancer cells within 10 h in serum-free medium. Scale bar: 200 μm.Real-time PCR showing that treatment of ME180 cells with AREG for 24 h significantly increased AREG mRNA expression. Each bar represents mean ± SEM (n = 9). Bars with different letters are significantly different from each other (P < 0.0001).Real-time PCR showing that knockdown of YAP in ME180 cells with YAP siRNA (siYAP) significantly suppressed AREG-induced AREG mRNA expression. siCTRL (non-target siRNA) was used as a siRNA control. Each bar represents mean ± SEM (n = 4). Bars with different letters are significantly different from each other (siCTRL vs. siCTRL+AREG, P < 0.0001; siYAP+CTRL vs. siYAP+AREG, P < 0.0001; siCTRL+AREG vs. siYAP+AREG, P = 0.0037).Data information: Quantitative data in (B) and (D) were analyzed for significance using unpaired t-test in GraphPad Prism 5 with Welch’s correction. Data in (E) were analyzed for significance using one-way ANOVA in GraphPad Prism 5 with Tukey’s post hoc tests. Source data are available online for this figure.

Mentions: We found that TGF-α treatment and YAP overexpression significantly increased AREG mRNA level (Fig6). Since AREG is a member of the family of the EGF-like ligands and we have shown that the EGFR pathway interacts with the Hippo pathway to regulate cervical cancer cell growth, we infer that AREG may also be involved in the regulation of cervical cancer cell proliferation. Treatment of ME180 cells with recombinant human AREG increased phosphorylation of EGFR at Tyr1173 and reduced phosphorylation of YAP (at Ser127 and Ser397), LATS1 (Ser 909), and MOB1 (Thr35) within 30 min (Fig7A, Appendix Fig S13). Treatment of ME180 cells with AREG induced elongated cell morphology (within 24 h) and significantly increased cell proliferation (72 h) (Fig7B). Moreover, AREG potently stimulated ME180 cell migration, as indicated by the significant increase in the wound closure in the wound-healing assay (Fig7C). Most interestingly, we found that the AREG mRNA expression was induced by AREG itself in cultured ME180 cervical cancer cells (Fig7D). Knockdown of YAP with YAP siRNA significantly suppressed AREG-stimulated AREG mRNA expression (P < 0.0001) (Fig7E).


The Hippo/YAP pathway interacts with EGFR signaling and HPV oncoproteins to regulate cervical cancer progression.

He C, Mao D, Hua G, Lv X, Chen X, Angeletti PC, Dong J, Remmenga SW, Rodabaugh KJ, Zhou J, Lambert PF, Yang P, Davis JS, Wang C - EMBO Mol Med (2015)

Function and expression of AREG in cervical cancer cellsWestern blot analysis showing that AREG treatment induced dephosphorylation of LATS1, MOB1, and YAP. ME180 cells were starved for 6 h after reaching cell confluence, then cells were treated with AREG (50 ng/ml) for 0, 30, 60 min.AREG (50 ng/ml) treatment induced appearance of elongated cells in ME180 cells (top and middle panels) and significantly stimulated ME180 cell proliferation (lower panel). Each bar represents mean ± SEM (n = 5). Bars with different letters are significantly different from each other (P = 0.003). Scale bar: 100 μm.Wound-healing assay showing that AREG stimulates migration of ME180 cervical cancer cells within 10 h in serum-free medium. Scale bar: 200 μm.Real-time PCR showing that treatment of ME180 cells with AREG for 24 h significantly increased AREG mRNA expression. Each bar represents mean ± SEM (n = 9). Bars with different letters are significantly different from each other (P < 0.0001).Real-time PCR showing that knockdown of YAP in ME180 cells with YAP siRNA (siYAP) significantly suppressed AREG-induced AREG mRNA expression. siCTRL (non-target siRNA) was used as a siRNA control. Each bar represents mean ± SEM (n = 4). Bars with different letters are significantly different from each other (siCTRL vs. siCTRL+AREG, P < 0.0001; siYAP+CTRL vs. siYAP+AREG, P < 0.0001; siCTRL+AREG vs. siYAP+AREG, P = 0.0037).Data information: Quantitative data in (B) and (D) were analyzed for significance using unpaired t-test in GraphPad Prism 5 with Welch’s correction. Data in (E) were analyzed for significance using one-way ANOVA in GraphPad Prism 5 with Tukey’s post hoc tests. Source data are available online for this figure.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4644376&req=5

fig07: Function and expression of AREG in cervical cancer cellsWestern blot analysis showing that AREG treatment induced dephosphorylation of LATS1, MOB1, and YAP. ME180 cells were starved for 6 h after reaching cell confluence, then cells were treated with AREG (50 ng/ml) for 0, 30, 60 min.AREG (50 ng/ml) treatment induced appearance of elongated cells in ME180 cells (top and middle panels) and significantly stimulated ME180 cell proliferation (lower panel). Each bar represents mean ± SEM (n = 5). Bars with different letters are significantly different from each other (P = 0.003). Scale bar: 100 μm.Wound-healing assay showing that AREG stimulates migration of ME180 cervical cancer cells within 10 h in serum-free medium. Scale bar: 200 μm.Real-time PCR showing that treatment of ME180 cells with AREG for 24 h significantly increased AREG mRNA expression. Each bar represents mean ± SEM (n = 9). Bars with different letters are significantly different from each other (P < 0.0001).Real-time PCR showing that knockdown of YAP in ME180 cells with YAP siRNA (siYAP) significantly suppressed AREG-induced AREG mRNA expression. siCTRL (non-target siRNA) was used as a siRNA control. Each bar represents mean ± SEM (n = 4). Bars with different letters are significantly different from each other (siCTRL vs. siCTRL+AREG, P < 0.0001; siYAP+CTRL vs. siYAP+AREG, P < 0.0001; siCTRL+AREG vs. siYAP+AREG, P = 0.0037).Data information: Quantitative data in (B) and (D) were analyzed for significance using unpaired t-test in GraphPad Prism 5 with Welch’s correction. Data in (E) were analyzed for significance using one-way ANOVA in GraphPad Prism 5 with Tukey’s post hoc tests. Source data are available online for this figure.
Mentions: We found that TGF-α treatment and YAP overexpression significantly increased AREG mRNA level (Fig6). Since AREG is a member of the family of the EGF-like ligands and we have shown that the EGFR pathway interacts with the Hippo pathway to regulate cervical cancer cell growth, we infer that AREG may also be involved in the regulation of cervical cancer cell proliferation. Treatment of ME180 cells with recombinant human AREG increased phosphorylation of EGFR at Tyr1173 and reduced phosphorylation of YAP (at Ser127 and Ser397), LATS1 (Ser 909), and MOB1 (Thr35) within 30 min (Fig7A, Appendix Fig S13). Treatment of ME180 cells with AREG induced elongated cell morphology (within 24 h) and significantly increased cell proliferation (72 h) (Fig7B). Moreover, AREG potently stimulated ME180 cell migration, as indicated by the significant increase in the wound closure in the wound-healing assay (Fig7C). Most interestingly, we found that the AREG mRNA expression was induced by AREG itself in cultured ME180 cervical cancer cells (Fig7D). Knockdown of YAP with YAP siRNA significantly suppressed AREG-stimulated AREG mRNA expression (P < 0.0001) (Fig7E).

Bottom Line: The Hippo signaling pathway controls organ size and tumorigenesis through a kinase cascade that inactivates Yes-associated protein (YAP).Our results suggest that YAP expression is associated with a poor prognosis for cervical cancer.Our study indicates that combined targeting of the Hippo and the ERBB signaling pathways represents a novel therapeutic strategy for prevention and treatment of cervical cancer.

View Article: PubMed Central - PubMed

Affiliation: Olson Center for Women's Health, Department of Obstetrics & Gynecology, University of Nebraska Medical Center, Omaha, NE, USA College of Animal Science and Technology, Huazhong Agricultural University, Wuhan, China.

Show MeSH
Related in: MedlinePlus