@article{16d38aa590dd4822a74e12e40c5b9be3,
title = "Depleting RhoA/stress fiber-organized fibronectin matrices on tumor cells non-autonomously aggravates fibroblast-driven tumor cell growth",
abstract = "Fibronectin (FN) expressed by tumor cells has been known to be tumor suppressive but the pericellular FN (periFN) assembled on circulating tumor cells appears to evidently promote distant metastasis. Whereas the regulation of periFN assembly in suspended cells has currently been under investigation, how it is regulated in adherent tumor cells and the role of periFN in primary tumor growth remain elusive. Techniques of RNAi, plasmid transfections, immunoblotting, fluorescence/immunohistochemistry staining, cell proliferation assays, and primary tumor growth in C57BL6 mice and Fischer 344 rats were employed in this study. We found that endogenously synthesized FN in adherent tumor cells was required for periFN assembly which was aligned by RhoA-organized actin stress fiber (SF). Depleting periFN on adherent tumor cells congruently promoted in vivo tumor growth but surprisingly did not autonomously impact on in vitro tumor cell proliferation and apoptosis, suggestive of a non-autonomous role of periFN in in vivo tumor growth. We showed that the proliferative ability of shFN-expressing tumor cells was higher than shScramble cells did in the presence of fibroblasts. Altogether, these results suggested that depriving RhoA/SF-regulated periFN matrices non-autonomously promotes fibroblast-mediated tumor cell growth.",
author = "Huang, {Li Tzu} and Tsai, {Chen Lung} and Huang, {Shin Huei} and Chang, {Ming Min} and Chang, {Wen Tsan} and Cheng, {Li Hsin} and Cheng, {Hung Chi}",
note = "Funding Information: All pharmaceutical compounds unless otherwise indicated and polyclonal antibodies (pAbs) against FN were purchased from Sigma-Aldrich, Inc. (St. Louis, MO, USA). 4′,6-diamidino-2-phenylindole (DAPI), Hoechst 33258, Phalloidin-Alexa 594 (A-12381), goat anti rabbit (GαR)-Alexa 488 (A11008), GαR-Alexa 594 (A11035) and pcDNA{\texttrademark}3.1/pcDNA-eGFP were from Invitrogen (Waltham, MA, USA). Rabbit pAb against α-smooth muscle actin (α-SMA) was from ProteinTech Group, Inc. (Chicago, IL, USA). Control rabbit non-immune IgG was from Jackson Immunoresearch Laboratories, INC (West Grove, PA, USA). The RNAi reagents for lentiviral vector system were from the National RNAi Core Facility supported by the National Research Program for Genomic Medicine Grants of NSC (NSC 100-2314-B-006-055). Bovine serum albumin (BSA), CyECL reagents and Cytochalasin D, an inhibitor of actin polymerization, were from Cyrusbioscience (Taipei, Taiwan). Annexin V-FAM apoptosis detection reagent was from LEADGENE (Tainan, Taiwan). TAlink mouse/rabbit polymer detection system (TAHC04D) was from BioTnA (Kaohsiung, Taiwan). Immobilon-P Poly vinylidene fluoride (PVDF) Membrane (IPVH00010) was from Sigma-Aldrich; Merck Millipore (Darmstadt, Germany). 12 mm microscope cover glasses were from Glaswarenfavrik Karl Hecht GmbH and Co. (Sondheim, Germany). Plasmids pRK5, p-RK5-RhoA-DN (RhoA-DN), and p-RK5-RhoA-CA (RhoA-CA) [81] were kindly received from Dr. A. Hall at MRC Laboratory for Molecular Cell Biology and Cell Biology Unit, London, UK. Funding Information: Funding: This work was supported by Ministry of Science and Technology, Taiwan (MOST106-2320-B-006-068-MY3 and MOST109-2320-B-006-022-).",
year = "2020",
month = nov,
day = "1",
doi = "10.3390/ijms21218272",
language = "English",
volume = "21",
pages = "1--20",
journal = "International Journal of Molecular Sciences",
issn = "1661-6596",
publisher = "Multidisciplinary Digital Publishing Institute (MDPI)",
number = "21",
}