br In conclusion the in vitro and in vivo results
In conclusion, the in vitro and in vivo results from the present study show that YAP1, the major effector of the Hippo pathway, plays a central role in the development of cervical cancer. Hyperactivation of YAP1 is sufficient to induce malignant trans-formation of cervical epithelial Necrosulfonamide and development of invasive cervical cancer in mouse models. Disruption of the Hippo pathway and subsequent activation of YAP1 in cervical epithelial cells may facilitate the establishment of persistent HPV infection by upregulating putative HPV receptor molecules and suppressing host cell innate immunity. Together with our previous observations that HPV16 E6 protein is able to suppress the Hippo pathway (thereby activating YAP1) and prevent YAP1 from proteasome-mediated degradation, we conclude that the interaction between the Hippo/YAP pathway hrHPV is a key player in cervical carcinogenesis. The present findings are ex-pected to influence current cervical cancer preventive screening, early detection, and treatment. The current HPV vaccination pro-gram may greatly reduce infection by HPV and carcinogenesis of the cervix in women in the near future. The novel mouse models developed in this study provide new tools for further investiga-tion of cervical cancer initiation and progression.
Detailed methods are provided in the online version of this paper and include the following:
d KEY RESOURCES TABLE
d CONTACT FOR REAGENT AND RESOURCE SHARING
d EXPERIMENTAL MODEL AND SUBJECT DETAILS B Mouse Model Studies
B Cell Culture
B Human Cervical Tissue Microarray
d METHOD DETAILS
B Reagents and Materials
B Fluorescent Immunocytochemistry B Western Blot Analysis
B Quantitative Real Time-PCR B YAP1 Overexpression
B Gene Silencing by siRNA B Preparation of HPV PsVs B HPV16 PsV Infection of Cells d QUANTIFICATION AND STATISTICAL ANALYSIS
Supplemental Information can be found with this article online at https://doi.
This work was supported by the National Cancer Institute (1R01CA197976 and 1R01CA201500); the Colleen’s Dream Foundation; the Olson Center for Women’s Health; the Fred & Pamela Buffett Cancer Center (LB595); the Marsha Rivkin Center for Cancer Research (the Barbara Learned Bridge Fund-ing Award); the Vincent Memorial Hospital Foundation; and the Vincent Center for Reproductive Biology.
C. He contributed to experimental design, performance, data analysis, and manuscript preparation. X.L. contributed to studies related to animal model establishment, data analysis, and manuscript preparation. C. Huang con-ducted IHC and immunofluorescence (IF) analyses and manuscript prepara-tion. G.H. packaged the retrovirus and performed some gene knockdown experiments. J.D. constructed YAP retroviral vectors. P.C.A. contributed to the construction of HPV16 E6/E7 expression vectors and establishment of the HPV16 PsVs. J.Z. contributed to histopathological analysis. B.M. and X.C. performed real-time PCR analysis. Z.W. contributed to western blotting analysis. P.F.L. contributed to the establishment of HPV-associated mouse models. B.R.R. contributed to manuscript preparation. J.S.D. contributed to data analysis and manuscript preparation. C.W. supervised these studies and contributed to experimental design, data analysis, and manuscript preparation.
DECLARATION OF INTERESTS
The authors declare no competing interests.
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