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| Status |
Public on Mar 08, 2017 |
| Title |
Shear stress activates YAP to promote motility in human prostate cancer cells |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
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| Summary |
Biophysical features of the microenvironment such as stiffness of extracellular matrix (ECM), nanotopography, and biomechanical force are critical regulators of cellular potential and behavior, yet the effects of extrinsic mechanical cues on tumor cells remain poorly understood. Here we demonstrate that frictional force, or wall shear stress (WSS), caused by fluid flow supports invasive behavior in cancer cells through activation of negative effectors of the Hippo tumor suppressor pathway, YAP and TAZ. In biomimetic models of lymphatic vasculature, WSS stimulated motility. These effects were accompanied by YAP dephosphorylation at ser-127, YAP and TAZ nuclear localization, and transactivation of YAP/TAZ downstream targets, including CTGF, AMOTL2, and ANKRD1. YAP, but not TAZ, was strictly required for WSS-enhanced motility, as knockdown of YAP or blockade of YAP-TEAD interactions by a small molecule inhibitor, verteporfin, reduced cellular velocity to levels observed in static controls. YAP-mediated effects on motility were dependent upon Rho-associated kinase (ROCK) and LIM-domain kinase (LIMK), as pharmacological inhibition of their activity led to activation of the actin-severing protein cofilin and blocked YAP dephosphorylation by WSS, thereby impairing migration. These data provide a signaling mechanism whereby biomechanical forces may influence cancer cell metastasis and implicate YAP as a core component of mechanosensitive machinery that modulates cancer progression.
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| Overall design |
The human prostate cancer cell line PC3 was tested for response to fluid wall shear stress (WSS). One day after seeding in a 6 well plate, cells of siRNA groups were transfected with control siRNA or YAP1 siRNA at 25nM using DharmaFect reagent. 24 hours after transfection, both tranfected cells and normal PC3 cells (10^5 cells per channel; surface area of each channel equaled 248 cm^2) were transferred into each polydimethylsiloxane (PDMS) channel (2.2 MPa surface stiffness) and permitted to attach for 24 hours. Fluid flow was then applied to each channel using a Harvard Apparatus PHD ULTRA programmable syringe pump. Cells were exposed either to static or WSS (0.05 dyne/cm^2) for 3 hours. Upon collection of cells with RLT lysis buffer (QIAGEN RNeasy kit), three channels of identical treatment were pooled to comprise a single sample. 12 samples total are included in this study. In detail, samples included at 3 hours: 3 static, 3 WSS, 3 WSS with siCTL and 3 WSS with siYAP. Sample labels end with the replicate number, i.e., WSS3 for the third replicate of the WSS sample.
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| Contributor(s) |
Wenzel PL |
| Citation(s) |
28098159 |
| NIH grant(s) |
| Grant ID |
Grant title |
Affiliation |
Name |
| K01 DK092365 |
Identification of biomechanical pathways that promote hematopoiesis |
UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON |
PAMELA LYNN WENZEL |
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| Submission date |
Sep 21, 2015 |
| Last update date |
Aug 13, 2018 |
| Contact name |
Pamela L Wenzel |
| Organization name |
University of Texas Medical School at Houston
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| Department |
Pediatric Surgery
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| Lab |
Pamela Wenzel
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| Street address |
1825 Pressler St, SRB637A
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| City |
Houston |
| State/province |
TX |
| ZIP/Postal code |
77030 |
| Country |
USA |
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| Platforms (1) |
| GPL10558 |
Illumina HumanHT-12 V4.0 expression beadchip |
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| Samples (12)
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| Relations |
| BioProject |
PRJNA296532 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE73284_RAW.tar |
51.0 Mb |
(http)(custom) |
TAR (of IDAT) |
| GSE73284_non-normalized.txt.gz |
2.3 Mb |
(ftp)(http) |
TXT |
| Raw data provided as supplementary file |
| Processed data included within Sample table |
| Raw data are available on Series record |
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