Page 25 - H. Cell signaling

Gallotannin regulates differentiation by changing cell morphology and Actin
cytoskeleton in Human osteoblast-like Saos-2 cells

Byung Su Ko, Yohan Han, Young Seok Eom and Song Ja Kim
Department of Biological Sciences, College of Natural Sciences, Kongju National University, Gongju, Chungnam 32588, Republic of Korea
BACKGROUND AIM

Osteosarcoma, one of the most common solid bone tumors in children and adolescents, often The effects of GT on
occurs in the metaphyseal region of the proximal humerus and proximal tibia. Though a differentiation in Human
combination of surgery, postoperative radiotherapy and chemotherapy is often used to treat osteoblast-like Saos-2 cells
osteosarcoma, osteosarcoma often has high morbidity and mortality. New methods for are not clearly understood.
osteosarcoma treatment are urgently needed. Several biological functions, such as cell So, we investigated effect of
viability, proliferation, migration, differentiation, tissue morphogenesis, and extracellular GT on differentiation
matrix (ECM) deposition are regulated by cell‐to‐matrix adhesion. To perform their function, changing of cell morphology
osteoblasts rely almost entirely on adhesion to the surrounding matrix. Gallotannin (GT) is a and actin cytoskeleton in
hydrolyzable tannin, i.e., a plant polyphenol, which is used as a protective agent in a similar Human osteoblast-like Saos-
manner to a drug, and exhibits strong activities against cancers, oxidation, viruses, bacteria, 2 cells.
and parasites.

METHODS
Cell culture Saos-2 osteoblastic cells, a human osteosarcoma osteoblastlike cells line (HTB-85; American Type Culture Collection), were cultured in McCoy’s 5A medium
supplemented with 100 U/ml penicillin/streptomycin, 15 % fetal bovine serum (FBS), at 37 °C and 5 % CO2 in a humidified incubator.
Western blot Thirty micrograms of protein samples were separated in SDS-polyacrylamide gel and transferred to a nitrocellulose membrane. Transfer was verified by Ponceau S
staining. The membrane was blocked for 60 min at room temperature with 5 % non-fat dry milk in T-TBS. Primary anti-COL2A1, anti-COL1A1, anti-Aggrecan, anti-GAPDH, anti-α-
tubulin and anti-β-actin diluted as indicated by manufacturers, were added overnight at 4 °C.
Immunofluorescence Revelation of α-tubulin, and ꞵ-actin within cells was identified. Secondary antibodies were incubated at room temperature for 2 h. Then, the cells were
counterstained with 4'6'-diamidino-2-phenylindole dihydrochloride (DAPI; Invitrogen; Thermo Fisher Scientific, Inc.). Fluorescent images were captured using a BX51 fluorescence
microscope (Olympus Corporation)
RESULTS

CONCLUSION REFERENCES Contact information

• We demonstrate that GT regulates 1. Mohamed Ibrahim, Julia Schoelermann, Kamal Mustafa and Mihaela R. Department of Biological
differentiation of in Human osteoblast-like Cimpan, TiO 2 nanoparticles disrupt cell adhesion and the architecture of Sciences, College of Natural
cytoskeletal networks of human osteoblast‐like cells in a size dependent
Saos-2 cells. manner. Journal of Biomedical Materials Research 106: 2575-2788, 2018 Sciences, Kongju National
• And, we verify that GT changes cell 2. Quanxiu Li, Guangchun Li, Changyi Liu, Na Chen, Bangyu Deng, and University, Gongju, Chungnam
morphology and actin cytoskeleton in Youke Xie, Cell Differentiation Agent-2 (CDA-2) Inhibits the Growth and 32588, Republic of Korea
Human osteoblast-like Saos-2 cells. Migration of Saos-2 Cells via miR-124/MAPK1. Cancer Management and
Research, 12: 4541-4548, 2020
• Therefore, we testify that GT regulates
differentiation by changing of cell 3. Su Min Kim, Yohan Han, Seon-Mi Yu and Song Ja Kim, Gallotannin Byung Su Ko and Song Ja Kim
attenuates 2-deoxy-D-glucose-induced dedifferentiation and endoplasmic
morphology and actin cytoskeleton in reticulum stress through inhibition of inositol-requiring enzyme 1
Human osteoblast-like Saos-2 cells. downstream p38 kinase pathway in chondrocytes. Molecular Medicine
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