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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.

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The Hippo/YAP and the ERBB pathways interact with each other to regulate cervical cancer cell growthRepresentative images showing the morphology of spheroids derived from ME180-siCtrl and ME180-siLATS1/2 cells growing in a 3D hanging-drop culture system for 10 days. Scale bar: 1.0 mm.Quantitative data showing changes in the volume of spheroids derived from ME180-siCTRL and ME180-siLATS1/2 cells growing in a 3D hanging-drop culture system. Each bar represents mean ± SEM (n = 5). Bars with different letters are significantly different from each other (P = 0.0004).Quantitative data showing changes in the volume of spheroids derived from ME180 cells growing in a 3D hanging-drop culture system in the absence or presence of AREG (20 ng/ml, 8 days). Each bar represents mean ± SEM (n = 5). Bars with different letters are significantly different from each other (P = 0.0030).Soft agar assay showing the effect of AG1478 and VTPF on colony formation in ME180-MXIV, ME180-YAP and ME180-YAPS127A cells. Scale bar: 500 μm.Representative images showing the effect of AREG, EGFR inhibitor (AG1478), and YAP antagonist verteporfin (VTPF) on the growth of ME180 cell in a 3D hanging-drop culture system. ME180 cells were incubated in the 3D hanging-drop culture system for 10 days in the absence or presence of AREG, AG1478 or verteporfin for 8 days. Scale bar: 1.0 mm.Data information: Quantitative data in (B) and (C) were analyzed for significance using unpaired t-test in GraphPad Prism 5 with Welch’s correction. Source data are available online for this figure.
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fig08: The Hippo/YAP and the ERBB pathways interact with each other to regulate cervical cancer cell growthRepresentative images showing the morphology of spheroids derived from ME180-siCtrl and ME180-siLATS1/2 cells growing in a 3D hanging-drop culture system for 10 days. Scale bar: 1.0 mm.Quantitative data showing changes in the volume of spheroids derived from ME180-siCTRL and ME180-siLATS1/2 cells growing in a 3D hanging-drop culture system. Each bar represents mean ± SEM (n = 5). Bars with different letters are significantly different from each other (P = 0.0004).Quantitative data showing changes in the volume of spheroids derived from ME180 cells growing in a 3D hanging-drop culture system in the absence or presence of AREG (20 ng/ml, 8 days). Each bar represents mean ± SEM (n = 5). Bars with different letters are significantly different from each other (P = 0.0030).Soft agar assay showing the effect of AG1478 and VTPF on colony formation in ME180-MXIV, ME180-YAP and ME180-YAPS127A cells. Scale bar: 500 μm.Representative images showing the effect of AREG, EGFR inhibitor (AG1478), and YAP antagonist verteporfin (VTPF) on the growth of ME180 cell in a 3D hanging-drop culture system. ME180 cells were incubated in the 3D hanging-drop culture system for 10 days in the absence or presence of AREG, AG1478 or verteporfin for 8 days. Scale bar: 1.0 mm.Data information: Quantitative data in (B) and (C) were analyzed for significance using unpaired t-test in GraphPad Prism 5 with Welch’s correction. Source data are available online for this figure.

Mentions: Treatment of confluent cervical cells with TGF-α and AREG resulted in a rapid and significant decrease in phosphorylation of LATS1, MOB1, and YAP (Figs6A and B and 7A, Appendix Figs S9, S10 and S13), suggesting that the Hippo pathway may involve in the YAP and EGFR signaling interaction. LATS1 and LATS2 are main components of the Hippo pathway and can directly phosphorylate YAP at Ser127. Knockdown of LATS1/2 in ME180 cells with LATS1/2 siRNAs activated YAP, which is indicated by a significant decrease in phospho-YAP (S127) (FigEV4A). Knockdown of LATS1/2 in ME180 cells also significantly increased cell proliferation and enhanced anchorage-independent cell growth (FigEV4B and C). The advantage of 3D culture, especially its high physiological relevance, has been reported (Friedrich et al, 2009). We found that knockdown of LATS1/2 significantly induced cell growth in the 3D culture system (Fig8A and B). Importantly, knockdown of LATS1/2 significantly increased the AREG secretion in both 2D and 3D culture (Fig EV4D and E). Consistent with 2D culture results, treatment of ME180 cells with AREG also significantly stimulated cell growth in the 3D culture system (Fig8C and E).


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)

The Hippo/YAP and the ERBB pathways interact with each other to regulate cervical cancer cell growthRepresentative images showing the morphology of spheroids derived from ME180-siCtrl and ME180-siLATS1/2 cells growing in a 3D hanging-drop culture system for 10 days. Scale bar: 1.0 mm.Quantitative data showing changes in the volume of spheroids derived from ME180-siCTRL and ME180-siLATS1/2 cells growing in a 3D hanging-drop culture system. Each bar represents mean ± SEM (n = 5). Bars with different letters are significantly different from each other (P = 0.0004).Quantitative data showing changes in the volume of spheroids derived from ME180 cells growing in a 3D hanging-drop culture system in the absence or presence of AREG (20 ng/ml, 8 days). Each bar represents mean ± SEM (n = 5). Bars with different letters are significantly different from each other (P = 0.0030).Soft agar assay showing the effect of AG1478 and VTPF on colony formation in ME180-MXIV, ME180-YAP and ME180-YAPS127A cells. Scale bar: 500 μm.Representative images showing the effect of AREG, EGFR inhibitor (AG1478), and YAP antagonist verteporfin (VTPF) on the growth of ME180 cell in a 3D hanging-drop culture system. ME180 cells were incubated in the 3D hanging-drop culture system for 10 days in the absence or presence of AREG, AG1478 or verteporfin for 8 days. Scale bar: 1.0 mm.Data information: Quantitative data in (B) and (C) were analyzed for significance using unpaired t-test in GraphPad Prism 5 with Welch’s correction. 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

fig08: The Hippo/YAP and the ERBB pathways interact with each other to regulate cervical cancer cell growthRepresentative images showing the morphology of spheroids derived from ME180-siCtrl and ME180-siLATS1/2 cells growing in a 3D hanging-drop culture system for 10 days. Scale bar: 1.0 mm.Quantitative data showing changes in the volume of spheroids derived from ME180-siCTRL and ME180-siLATS1/2 cells growing in a 3D hanging-drop culture system. Each bar represents mean ± SEM (n = 5). Bars with different letters are significantly different from each other (P = 0.0004).Quantitative data showing changes in the volume of spheroids derived from ME180 cells growing in a 3D hanging-drop culture system in the absence or presence of AREG (20 ng/ml, 8 days). Each bar represents mean ± SEM (n = 5). Bars with different letters are significantly different from each other (P = 0.0030).Soft agar assay showing the effect of AG1478 and VTPF on colony formation in ME180-MXIV, ME180-YAP and ME180-YAPS127A cells. Scale bar: 500 μm.Representative images showing the effect of AREG, EGFR inhibitor (AG1478), and YAP antagonist verteporfin (VTPF) on the growth of ME180 cell in a 3D hanging-drop culture system. ME180 cells were incubated in the 3D hanging-drop culture system for 10 days in the absence or presence of AREG, AG1478 or verteporfin for 8 days. Scale bar: 1.0 mm.Data information: Quantitative data in (B) and (C) were analyzed for significance using unpaired t-test in GraphPad Prism 5 with Welch’s correction. Source data are available online for this figure.
Mentions: Treatment of confluent cervical cells with TGF-α and AREG resulted in a rapid and significant decrease in phosphorylation of LATS1, MOB1, and YAP (Figs6A and B and 7A, Appendix Figs S9, S10 and S13), suggesting that the Hippo pathway may involve in the YAP and EGFR signaling interaction. LATS1 and LATS2 are main components of the Hippo pathway and can directly phosphorylate YAP at Ser127. Knockdown of LATS1/2 in ME180 cells with LATS1/2 siRNAs activated YAP, which is indicated by a significant decrease in phospho-YAP (S127) (FigEV4A). Knockdown of LATS1/2 in ME180 cells also significantly increased cell proliferation and enhanced anchorage-independent cell growth (FigEV4B and C). The advantage of 3D culture, especially its high physiological relevance, has been reported (Friedrich et al, 2009). We found that knockdown of LATS1/2 significantly induced cell growth in the 3D culture system (Fig8A and B). Importantly, knockdown of LATS1/2 significantly increased the AREG secretion in both 2D and 3D culture (Fig EV4D and E). Consistent with 2D culture results, treatment of ME180 cells with AREG also significantly stimulated cell growth in the 3D culture system (Fig8C and E).

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