Differential Contributions of Actin and Myosin to the Physical Phenotypes and Invasion of Pancreatic Cancer Cells
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Metastasis is a fundamentally physical process in which cells deform through narrow gaps and generate forces to invade surrounding tissues. While it is commonly thought that increased cell deformability is an advantage for invading cells, we previously found that more invasive pancreatic ductal adenocarcinoma (PDAC) cells are stiffer than less invasive PDAC cells. Here we investigate potential mechanisms of the simultaneous increase in PDAC cell stiffness and invasion, focusing on the contributions of myosin II, Arp2/3, and formins.
We measure cell invasion using a 3D scratch wound invasion assay and cell stiffness using atomic force microscopy (AFM). To determine the effects of actin- and myosin-mediated force generation on cell stiffness and invasion, we treat cells with pharmacologic inhibitors of myosin II (blebbistatin), Arp2/3 (CK-666), and formins (SMIFH2).
We find that the activity of myosin II, Arp2/3, and formins all contribute to the stiffness of PDAC cells. Interestingly, we find that the invasion of PDAC cell lines is differentially affected when the activity of myosin II, Arp2/3, or formins is inhibited, suggesting that despite having similar tissue origins, different PDAC cell lines may rely on different mechanisms for invasion.
These findings deepen our knowledge of the factors that regulate cancer cell mechanotype and invasion, and incite further studies to develop therapeutics that target multiple mechanisms of invasion for improved clinical benefit.
KeywordsMechanobiology Cytoskeleton Pancreatic ductal adenocarcinoma Cell stiffness Arp2/3 Formins Traction forces Cell motility
We thank our funding sources: the National Science Foundation (CAREER DBI-1254185 and BMMB-1906165 to ACR), the Farber Family Foundation, and UCLA Integrative Biology & Physiology Eureka Scholarship (to AVN), and the National Institutes of Health (R01 GM110482 to MJB). We would also like to thank Timothy Donahue and his laboratory for their insights into PDAC, as well as their generous contributions of the PDAC cell lines used in our studies. We are also grateful to Gordon Robertson and Ewan Gibb for their bioinformatics expertise. The MMP activity assay was performed in the UCLA Molecular Shared Screening Resource in the California NanoSystems Institute with technical support from Robert Damoiseaux and Bobby Tofig.
Conflict of interest
Angelyn V. Nguyen, Brittany Trompetto, Xing Haw Marvin Tan, Michael B. Scott, Kenneth Hsueh-heng Hu, Eric Deeds, Manish J. Butte, Pei Yu Chiou, and Amy C. Rowat have no conflicts of interest.
No human or animals studies were carried out by the authors for this article.
- 22.Ellerbroek, S. M., et al. Ovarian carcinoma regulation of matrix metalloproteinase-2 and membrane type 1 matrix metalloproteinase through beta1 integrin. Cancer Res. 59:1635–1641, 1999.Google Scholar
- 37.Jimenez Valencia, A. M., et al. Collective cancer cell invasion induced by coordinated contractile stresses. Oncotarget 6:43438–43451, 2015.Google Scholar
- 62.Noël, A. C., et al. Invasion of reconstituted basement membrane matrix is not correlated to the malignant metastatic cell phenotype. Cancer Res. 51:405–414, 1991.Google Scholar
- 73.Rasheed, Z. A., W. Matsui, and A. Maitra. Pathology of pancreatic stroma in PDAC. In: Pancreatic Cancer and Tumor Microenvironment, edited by P. J. Grippo, and H. G. Munshi. Trivandrum: Transworld Research Network, 2012.Google Scholar
- 75.Rauhala, H. E., S. Teppo, S. Niemelä, and A. Kallioniemi. Silencing of the ARP2/3 complex disturbs pancreatic cancer cell migration. Anticancer Res. 33:45–52, 2013.Google Scholar
- 86.Shields, M. A., S. Dangi-Garimella, S. B. Krantz, D. J. Bentrem, and H. G. Munshi. Pancreatic cancer cells respond to type I collagen by inducing snail expression to promote membrane type 1 matrix metalloproteinase-dependent collagen invasion. J. Biol. Chem. 286:10495–10504, 2011.CrossRefGoogle Scholar
- 110.Xiao, F., X. Wen, and P. Y. Chiou. Plasmonic micropillars for massively parallel precision cell force measurement. Micro Electro Mech. Syst. 1:243–246, 2017.Google Scholar