TY - JOUR
T1 - RAS specific protease induces irreversible growth arrest via p27 in several KRAS mutant colorectal cancer cell lines
AU - Stubbs, Caleb K.
AU - Biancucci, Marco
AU - Vidimar, Vania
AU - Satchell, Karla J.F.
N1 - Funding Information:
We thank the Frederick National Laboratory for Cancer Research (FNLCR) for providing the RAS-less MEF cells, KRAS mutant cell lines, and hybridoma cells for the pan-RAS antibody directed against the G-domain. Matthew Kieffer, Megan Packer, and all members of the Satchell laboratory are thanked for valuable intellectual input and technical support. This work was funded by grants from the Chicago Biomedical Consortium, H Foundation, and Northwestern Medicine Catalyst Fund (to K.J.F.S). C.K.S. was supported by a fellowship from the National Cancer Institute (T32 CA09560). High content imaging was performed on the Nikon Biostation CT system purchased with the support of NIH grant S10 OD021704. Flow cytometry core services were provided by the Northwestern University RHLCCC Flow Cytometry Facility supported by NIH grant P30 CA060553.
Publisher Copyright:
© 2021, The Author(s).
PY - 2021/12
Y1 - 2021/12
N2 - Ras-specific proteases to degrade RAS within cancer cells are under active development as an innovative strategy to treat tumorigenesis. The naturally occurring biological toxin effector called RAS/RAP1-specific endopeptidase (RRSP) is known to cleave all RAS within a cell, including HRAS, KRAS, NRAS and mutant KRAS G13D. Yet, our understanding of the mechanisms by which RRSP drives growth inhibition are unknown. Here, we demonstrate, using isogenic mouse fibroblasts expressing a single isoform of RAS or mutant KRAS, that RRSP equally inactivates all isoforms of RAS as well as the major oncogenic KRAS mutants. To investigate how RAS processing might lead to varying outcomes in cell fate within cancer cells, we tested RRSP against four colorectal cancer cell lines with a range of cell fates. While cell lines highly susceptible to RRSP (HCT116 and SW1463) undergo apoptosis, RRSP treatment of GP5d and SW620 cells induces G1 cell cycle arrest. In some cell lines, growth effects were dictated by rescued expression of the tumor suppressor protein p27 (Kip1). The ability of RRSP to irreversibly inhibit cancer cell growth highlights the antitumor potential of RRSP, and further warrants investigation as a potential anti-tumor therapeutic.
AB - Ras-specific proteases to degrade RAS within cancer cells are under active development as an innovative strategy to treat tumorigenesis. The naturally occurring biological toxin effector called RAS/RAP1-specific endopeptidase (RRSP) is known to cleave all RAS within a cell, including HRAS, KRAS, NRAS and mutant KRAS G13D. Yet, our understanding of the mechanisms by which RRSP drives growth inhibition are unknown. Here, we demonstrate, using isogenic mouse fibroblasts expressing a single isoform of RAS or mutant KRAS, that RRSP equally inactivates all isoforms of RAS as well as the major oncogenic KRAS mutants. To investigate how RAS processing might lead to varying outcomes in cell fate within cancer cells, we tested RRSP against four colorectal cancer cell lines with a range of cell fates. While cell lines highly susceptible to RRSP (HCT116 and SW1463) undergo apoptosis, RRSP treatment of GP5d and SW620 cells induces G1 cell cycle arrest. In some cell lines, growth effects were dictated by rescued expression of the tumor suppressor protein p27 (Kip1). The ability of RRSP to irreversibly inhibit cancer cell growth highlights the antitumor potential of RRSP, and further warrants investigation as a potential anti-tumor therapeutic.
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U2 - 10.1038/s41598-021-97422-0
DO - 10.1038/s41598-021-97422-0
M3 - Article
C2 - 34504197
AN - SCOPUS:85115034570
SN - 2045-2322
VL - 11
JO - Scientific reports
JF - Scientific reports
IS - 1
M1 - 17925
ER -
