Comparison of Selected Non-Coding RNAs and Gene Expression Profiles between Common Osteosarcoma Cell Lines

Mateusz Sikora; Katarzyna Krajewska; Klaudia Marcinkowska; Anna Raciborska; Rafał Jakub Wiglusz; Agnieszka Śmieszek

doi:10.3390/cancers14184533

Comparison of Selected Non-Coding RNAs and Gene Expression Profiles between Common Osteosarcoma Cell Lines

Cancers (Basel). 2022 Sep 19;14(18):4533. doi: 10.3390/cancers14184533.

Authors

Mateusz Sikora¹, Katarzyna Krajewska¹, Klaudia Marcinkowska¹, Anna Raciborska², Rafał Jakub Wiglusz^{3

4

5}, Agnieszka Śmieszek¹

Affiliations

¹ The Department of Experimental Biology, The Faculty of Biology and Animal Science, University of Environmental and Life Sciences Wroclaw, Norwida 27B, 50-375 Wroclaw, Poland.
² Department of Oncology and Surgical Oncology for Children and Youth, Institute of Mother and Child, ul. Kasprzaka 17a, 01-211 Warsaw, Poland.
³ Institute of Low Temperature and Structure Research, Polish Academy of Sciences, Okolna 2, 50-422 Wroclaw, Poland.
⁴ Centre for Advanced Materials and Smart Structures, Polish Academy of Sciences, Okolna 2, 50-950 Wroclaw, Poland.
⁵ International Institute of Translational Medicine, Jesionowa 11 St, 55-124 Malin, Poland.

Abstract

Osteosarcoma (OS) is a bone tumour affecting adolescents and elderly people. Unfortunately, basic treatment methods are still underdeveloped, which has a high impact on the poor survivability of the patients. Studies designed to understand the underlying mechanisms of osteosarcoma development, as well as preclinical investigations aimed at establishing novel therapeutic strategies, rely significantly upon in vitro models, which apply well-established cell lines such as U-2 OS, Saos-2 and MG-63. In this study, the expression of chosen markers associated with tumour progression, metastasis and survival were identified using RT-qPCR. Levels of several onco-miRs (miR-21-5p, miR-124-3p, miR-223-3p and miR-320a-3p) and long non-coding RNA MEG3 were established. The mRNA expression of bone morphogenetic proteins (BMPs), including BMP-2, BMP-3, BMP-4, BMP-6, BMP-7, as well as their receptors: BMPR-IA, BMPR-IB and BMPR-II was also determined. Other tested markers included metalloproteinases, i.e., MMP-7 and MMP-14 and survivin (BIRC5), C-MYC, as well as CYCLIN D (CCND1). The analysis included comparing obtained profiles with transcript levels established for the osteogenic HeLa cell line and human adipose-derived stromal cells (hASCs). The tested OS cell lines were characterised by a cancer-related phenotype, such as increased expression of mRNA for BMP-7, as well as MMP-7 and MMP-14. Osteosarcoma cells differ considerably in miR-21-5p and miR-124-3p levels, which can be related to uncontrolled tumour growth. The comprehensive examination of osteosarcoma transcriptome profiles may facilitate the selection of appropriate cell models for preclinical investigations aimed at the development of new strategies for OS treatment.

Keywords: RT-qPCR; gene expression profile; mRNA; microRNA; osteosarcoma.

Grants and funding

The research is financed under the individual student research project “Młode umysły—Young Minds Project” from the subsidy increased for the period 2020–2026 in the amount of 2% of the subsidy referred to Art. 387 (3) of the Law of 20 July 2018 on Higher Education and Science, obtained in 2019. The publication is co-financed by project founded by Wroclaw University of Environmental and Life Science, i.e., “Bon doktoranta SD UPWr” no. N020/0017/20. Additional founding for project realization was supported by Medical Research Agency (ABM) under the project REGBONE (2021/ABM/01/00019).