Externally added cystatin C reduces growth of A375 melanoma cells by increasing cell cycle time
(2021) In FEBS Open Bio 11(6). p.1645-1658- Abstract
Some secreted cysteine protease inhibitors of the cystatin family appear to affect intracellular proteolysis and growth of human cells, as a result of internalization. Here, we studied the effects of external addition of the most abundant human cystatin, cystatin C, on viability and proliferation of cancer cells in culture. A dose-dependent decrease in viable cells was seen for A375 melanoma, MCF-7 breast cancer, and PC-3 prostate cancer cells cultured in 1–5 µm cystatin C after 24 h. Real-time assessment of growth rates in A375 cell cultures for 48 h by digital holographic microscopy showed an increased doubling time for cells cultured in the presence of 5 µm cystatin C (20.1 h) compared with control cells (14.7 h). A prolonged... (More)
Some secreted cysteine protease inhibitors of the cystatin family appear to affect intracellular proteolysis and growth of human cells, as a result of internalization. Here, we studied the effects of external addition of the most abundant human cystatin, cystatin C, on viability and proliferation of cancer cells in culture. A dose-dependent decrease in viable cells was seen for A375 melanoma, MCF-7 breast cancer, and PC-3 prostate cancer cells cultured in 1–5 µm cystatin C after 24 h. Real-time assessment of growth rates in A375 cell cultures for 48 h by digital holographic microscopy showed an increased doubling time for cells cultured in the presence of 5 µm cystatin C (20.1 h) compared with control cells (14.7 h). A prolonged doubling time was already observed during the first 12 h, indicating a rapid general decrease in cell proliferation at the population level. Tracking of individual cells in phase holographic images showed that dividing cells incubated with 5 µm cystatin C underwent fewer mitoses during 48 h than control cells. In addition, the time between cell divisions was longer, especially for the first cell cycle. Incubation with the variant W106F-cystatin C (with high cellular uptake rate) resulted in a lower number of viable cells and a prolonged doubling time than when cells were incubated with wild-type cystatin C, but no effect was observed for (R24A,R25A)-cystatin C (low cellular uptake). Thus, cystatin C causes prolonged cell division leading to decreased proliferation of melanoma cells, and internalization seems to be a prerequisite for this effect.
(Less)
- author
- Wallin, Hanna LU ; Hunaiti, Samar LU and Abrahamson, Magnus LU
- organization
- publishing date
- 2021-06
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- A375 cells, cell cycle, cell growth, cysteine peptidase, digital holographic microscopy, protease inhibitor
- in
- FEBS Open Bio
- volume
- 11
- issue
- 6
- pages
- 14 pages
- publisher
- Wiley-Blackwell
- external identifiers
-
- pmid:33837649
- scopus:85107251735
- ISSN
- 2211-5463
- DOI
- 10.1002/2211-5463.13162
- language
- English
- LU publication?
- yes
- id
- 94937dd1-1703-4776-ab7b-c19909825c75
- date added to LUP
- 2021-06-22 15:09:55
- date last changed
- 2025-01-12 10:31:45
@article{94937dd1-1703-4776-ab7b-c19909825c75,
abstract = {{<p>Some secreted cysteine protease inhibitors of the cystatin family appear to affect intracellular proteolysis and growth of human cells, as a result of internalization. Here, we studied the effects of external addition of the most abundant human cystatin, cystatin C, on viability and proliferation of cancer cells in culture. A dose-dependent decrease in viable cells was seen for A375 melanoma, MCF-7 breast cancer, and PC-3 prostate cancer cells cultured in 1–5 µm cystatin C after 24 h. Real-time assessment of growth rates in A375 cell cultures for 48 h by digital holographic microscopy showed an increased doubling time for cells cultured in the presence of 5 µm cystatin C (20.1 h) compared with control cells (14.7 h). A prolonged doubling time was already observed during the first 12 h, indicating a rapid general decrease in cell proliferation at the population level. Tracking of individual cells in phase holographic images showed that dividing cells incubated with 5 µm cystatin C underwent fewer mitoses during 48 h than control cells. In addition, the time between cell divisions was longer, especially for the first cell cycle. Incubation with the variant W106F-cystatin C (with high cellular uptake rate) resulted in a lower number of viable cells and a prolonged doubling time than when cells were incubated with wild-type cystatin C, but no effect was observed for (R24A,R25A)-cystatin C (low cellular uptake). Thus, cystatin C causes prolonged cell division leading to decreased proliferation of melanoma cells, and internalization seems to be a prerequisite for this effect.</p>}},
author = {{Wallin, Hanna and Hunaiti, Samar and Abrahamson, Magnus}},
issn = {{2211-5463}},
keywords = {{A375 cells; cell cycle; cell growth; cysteine peptidase; digital holographic microscopy; protease inhibitor}},
language = {{eng}},
number = {{6}},
pages = {{1645--1658}},
publisher = {{Wiley-Blackwell}},
series = {{FEBS Open Bio}},
title = {{Externally added cystatin C reduces growth of A375 melanoma cells by increasing cell cycle time}},
url = {{http://dx.doi.org/10.1002/2211-5463.13162}},
doi = {{10.1002/2211-5463.13162}},
volume = {{11}},
year = {{2021}},
}