Mutant p53 regulates ovarian cancer transformed phenotypes through autocrine matrix deposition
Marcin P. Iwanicki, … , Ronny Drapkin, Joan S. Brugge
Marcin P. Iwanicki, … , Ronny Drapkin, Joan S. Brugge
Published July 7, 2016
Citation Information: JCI Insight. 2016;1(10):e86829. https://doi.org/10.1172/jci.insight.86829.
View: Text | PDF
Research Article Cell biology Oncology

Mutant p53 regulates ovarian cancer transformed phenotypes through autocrine matrix deposition

Abstract

High-grade serous ovarian carcinoma (HGS-OvCa) harbors p53 mutations and can originate from the epithelial cell compartment of the fallopian tube fimbriae. From this site, neoplastic cells detach, survive in the peritoneal cavity, and form cellular clusters that intercalate into the mesothelium to form ovarian and peritoneal masses. To examine the contribution of mutant p53 to phenotypic alterations associated with HGS-OvCA, we developed live-cell microscopy assays that recapitulate these early events in cultured fallopian tube nonciliated epithelial (FNE) cells. Expression of stabilizing mutant variants of p53, but not depletion of endogenous wild-type p53, in FNE cells promoted survival and cell-cell aggregation under conditions of cell detachment, leading to the formation of cell clusters with mesothelium-intercalation capacity. Mutant p53R175H-induced phenotypes were dependent on fibronectin production, α5β1 fibronectin receptor engagement, and TWIST1 expression. These results indicate that FNE cells expressing stabilizing p53 mutants acquire anchorage independence and subsequent mesothelial intercalation capacity through a mechanism involving mesenchymal transition and matrix production. These findings provide important new insights into activities of mutant p53 in the cells of origin of HGS-OvCa.

Authors

Marcin P. Iwanicki, Hsing-Yu Chen, Claudia Iavarone, Ioannis K. Zervantonakis, Taru Muranen, Marián Novak, Tan A. Ince, Ronny Drapkin, Joan S. Brugge

×

Figure 8

Reversion of mutant p53R175H (m-p53R175H) conformation with zinc metallochaperone 1 (ZMC1) blocks m-p53R175H–induced phenotypes.

Options: View larger image (or click on image) Download as PowerPoint
Reversion of mutant p53R175H (m-p53R175H) conformation with zinc metallo...
(A) Representative phase-contrast video clips of DF30 cells treated with either DMSO or 250 nM ZMC1 and cultured in suspension for 8 hours (Supplemental Video 13). Representative images from 8 or 9 movies acquired per condition during 1 recording session. Scale bar: 100 μm. (B) Distribution of the percentage area reduction of DF30 cell clusters from the experiment in panel A. n = 8 or 9 cell clusters scored per condition. (C) Phase-contrast and pseudocolored (red) images showing the ability of DMSO- or 250 nM ZMC1–treated DF30 cells to clear the mesothelium at the indicated time points (Supplemental Video 14). Scale bar: 150 μm. (D) Quantification of the mesothelial clearance distribution by DF30 cells treated with DMSO or ZMC1 (250 nM). This experiment was repeated twice with n = 10 (DMSO) and n = 20 (ZMC1) cell clusters scored per experiment. (E) TWIST1 mRNA fold change in various fallopian tube nonciliated epithelial (FNE) cells. Values are normalized to FNE control cells. This experiment was performed 3 times with 3 technical replicates. (F) Representative phase-contrast movie clips of DF30 cells treated with either DMSO or 250 nM ZMC1 and cultured in suspension for the indicated time points (Supplemental Video 15). Arrows point to dying cells. (G) Pseudocolored fluorescence images showing ethidium bromide (EtBr) incorporation (red) by DF30 clusters (green) treated with DMSO or 250 nM ZMC1. (H) Distribution of EtBr incorporation by DF30 clusters from 3 independent experiments with n = 44 (DMSO) and n = 46 (ZMC1) clusters scored. Each cluster consisted of 100–150 cells. (I) Model of mutant p53–dependent survival of FNE cells that have exfoliated from fimbriae and intercalated into a mesothelial monolayer. All data shown as the median (horizontal bar), interquartile range (box), and minimum/maximum values (whiskers). Statistical analysis performed using 2-tailed Student’s t test. *P < 0.05.