A bulky glycocalyx fosters metastasis formation by promoting G1 cell cycle progression
- Stanford University, United States
- University of California, San Francisco, United States
- Howard Hughes Medical Institute, Stanford University, United States
Figures
Figure 1 with 3 supplements
The glycocalyx increases metastatic potential in a size dependent manner.
(A) Mucin-mimetic glycopolymers consist of a poly(methyl vinyl ketone) polymer with pendant GalNAc residues. The glycopolymer terminates in a synthetic sterol for insertion into cell membranes. …
Figure 1—figure supplement 1
Long glycopolymers delay MCF-10A cell death in nude mice.
MCF-10A cells were painted with either short or long glycopolymers and injected into the tail veins of nude mice. At the time points indicated, the mice were sacrificed and their lungs fixed and …
Figure 1—figure supplement 2
Long glycopolymer-bearing 4TO7 cells proliferate more rapidly in mouse lungs than those bearing short glycopolymers.
Animals were injected on day 0 with 4TO7 cells treated with either long or short cell-surface glycopolymers. At a given day post injection, mice were anesthetized, administered luciferin by …
Figure 1—figure supplement 3
Short glycopolymer-bearing 4TO7 cells have no metastatic advantage over naïve 4TO7 cells.
Animals were injected on day 0 with cells treated with either short cell-surface glycopolymers or PBS vehicle. (A) Quantification of bioluminescence imaging of anesthetized mice. At a given day post …
Figure 2 with 5 supplements
A thick glycocalyx drives cell cycle progression via the PI3K-Akt axis.
(A) and (B) Microscopy of MCF-10A cells labeled with short or long glycopolymers, plated on soft (400 Pa) fibronectin-functionalized gels, and cultured for 72 hr. (C) Average number of cells per …
Figure 2—figure supplement 1
Glycopolymers must be cell-surface resident to affect colony size.
(A) Average number of cells per colony of MCF-10A cells treated with cell-surface persistent (Long) or rapidly endocytosed but equally long (DPPE) mucin-mimetic glycopolymers then plated at low …
Figure 2—figure supplement 2
Glycopolymers do not abolish need for growth factor signals in proliferation.
(A) Immunoblot of MCF-10A cells serum-starved for 72 hr, treated with long or short glycopolymers or vehicle and plated in serum-free media on soft or stiff gels for six hours then lysed. Shown is a …
Figure 2—figure supplement 3
Long glycopolymers increase active Akt in 4TO7 lung metastases.
Tissue sections from lungs derived from the experiments described in Figure 1 were stained with an anti-pAkt substrate antibody. The antibody is selective for a phosphorylated consensus sequence …
Figure 2—figure supplement 4
PI3K or MEK inhibitor abolishes effects of long glycopolymers on proliferation.
(A) Average number of cells per colony of long glycopolymer-bearing MCF-10A cells plated on soft gels in media containing 50 μM PI3K inhibitor LY294002 or vehicle (DMSO) and cultured for 72 hr. (B) …
Figure 2—figure supplement 5
Cyclin D1 expression in 4TO7 cells in vitro.
A)Immunoblot analysis of Cyclin D1 levels in 4TO7 cells coated with long or short glycopolymers or treated with vehicle (PBS) and plated on soft (400 Pa) fibronectin-functionalized polyacrylamide …
Figure 3 with 1 supplement
A thick glycocalyx stimulates integrin-FAK mechanosignaling.
(A) Immunoblot analysis of FAK phosphorylation in MCF-10A cells coated with long or short glycopolymers or treated with vehicle (PBS) and plated on soft (400 Pa) or stiff (60 kPa) …
Figure 3—figure supplement 1
FAK inhibitor abolishes proliferative advantage from bulky glycocalyx.
(A) Average number of cells per colony of long glycopolymer-bearing MCF-10A cells plated on soft gels in media containing 1 μM FAK inhibitor FAK14 or vehicle (DMSO) and cultured for 72 hr. *p<0.05 …
Figure 4 with 2 supplements
The MUC1 ectodomain is sufficient to increase the metastatic potential of 4TO7 cancer cells.
(A) Experimental scheme for Figure 4. Balb/c mice were injected with syngeneic 4TO7 mammary carcinoma cells transfected with a MUC1ΔCT construct under the control of a doxycycline (dox)-inducible …
Figure 4—figure supplement 1
4TO7 cells expressing the MUC1 ectodomain proliferate more rapidly in mouse lungs than those that do not.
Animals were injected on day 0 with 4TO7 cells treated with either doxycycline to induce expression of MUC1ΔCT construct or PBS vehicle (Native). At a given day post injection, mice were …
Figure 4—figure supplement 2
The MUC1 ectodomain increases Akt activity in 4TO7 lung metastases.
Tissue sections from lungs derived from the experiments described in Figure 4 were stained with an anti-pAkt substrate antibody. The antibody is selective for a phosphorylated consensus sequence …
Author response image 1
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- https://doi.org/10.7554/eLife.25752.017
