Diamond Nanofilm Normalizes Proliferation and Metabolism in Liver Cancer Cells

Abstract

Malwina Sosnowska,1 Marta Kutwin,1 Barbara Strojny,1 Mateusz Wierzbicki,1 Dominik Cysewski,2 Jarosław Szczepaniak,1 Mateusz Ficek,3 Piotr Koczoń,4 Sławomir Jaworski,1 André Chwalibog,5 Ewa Sawosz1 1Department of Nanobiotechnology, Institute of Biology, Warsaw University of Life Sciences, Warsaw, Poland; 2Spectrometry Laboratory, Institute of Biochemistry and Biophysics, Polish Academy of Science, Warsaw, Poland; 3Department of Metrology and Optoelectronics, Gdansk University of Technology, Gdansk, Poland; 4Department of Chemistry, Institute of Food Sciences, Warsaw University of Life Sciences, Warsaw, Poland; 5Department of Veterinary and Animal, Sciences, University of Copenhagen, Frederiksberg, DenmarkCorrespondence: André ChwalibogDepartment of Veterinary and Animal Sciences, University of Copenhagen, Groennegaardsvej 3, Frederiksberg, 1870, DenmarkTel [phone]Email [email protected] SosnowskaDepartment of Nanobiotechnology, Institute of Biology, Warsaw University of Life Sciences, Warsaw, PolandTel [phone]Email [email protected]: Surgical resection of hepatocellular carcinoma can be associated with recurrence resulting from the degeneration of residual volume of the liver. The objective was to assess the possibility of using a biocompatible nanofilm, made of a colloid of diamond nanoparticles (nfND), to fill the side after tumour resection and optimize its contact with proliferating liver cells, minimizing their cancerous transformation.Methods: HepG2 and C3A liver cancer cells and HS-5 non-cancer cells were used. An aqueous colloid of diamond nanoparticles, which covered the cell culture plate, was used to create the nanofilm. The roughness of the resulting nanofilm was measured by atomic force microscopy. Mitochondrial activity and cell proliferation were measured by XTT and BrdU assays. Cell morphology and a scratch test were used to evaluate the invasiveness of cells. Flow cytometry determined the number of cells within the cell cycle. Protein expression in was measured by mass spectrometry.Results: The nfND created a surface with increased roughness and exposed oxygen groups compared with a standard plate. All cell lines were prone to settling on the nanofilm, but cancer cells formed more relaxed clusters. The surface compatibility was dependent on the cell type and decreased in the order C3A >HepG2 >HS-5. The invasion was reduced in cancer lines with the greatest effect on the C3A line, reducing proliferation and increasing the G2/M cell population. Among the proteins with altered expression, membrane and nuclear proteins dominated.Conclusion: In vitro studies demonstrated the antiproliferative properties of nfND against C3A liver cancer cells. At the same time, the need to personalize potential therapy was indicated due to the differential protein synthetic responses in C3A vs HepG2 cells. We documented that nfND is a source of signals capable of normalizing the expression of many intracellular proteins involved in the transformation to non-cancerous cells.Keywords: cell cycle, cell proteome, diamond nanofilm, extracellular matrix, invasion, liver cancer

Keywords

• cell cycle
• cell proteome
• diamond nanofilm
• extracellular matrix
• invasion
• liver cancer