A Comparative Study of Nucleostemin Gene Expression in Two-Dimensional Culture Medium and Multicellular Breast Cancer Spheroids

Subject Areas : Journal of Animal Biology

N. Rezapour ¹ , Vajiheh zarrinpour ² , mohammad kamalabadi farahani ³ , amir atashi ⁴

1 - Department of Biology, Damghan Branch, Islamic Azad University, Damghan, Iran
2 - Department of Biology, Damghan Branch, Islamic Azad University, Damghan, Iran
3 - Department of Tissue Engineering, School of Medicine, Shahroud University of Medical Sciences, Shahroud, Iran
4 - Department of Medical Laboratory Sciences, School of Paramedical, Shahroud University of Medical Sciences, Shahroud, Iran

Received: 2024-01-19 Accepted : 2024-02-04 Published : 2024-06-01

Keywords: Breast Cancer, Nucleostemin, Metastasis, Cancer Stem Cells,

Abstract :

Breast cancer mortality is mainly due to metastatic disease caused by cancer stem cells. Nucleostemin is a GTP-bound nuclear cofactor that is highly expressed in normal stem cells and tumors and is thought to play an important role in the pathogenesis and metastasis of breast cancer. This study examines the expression level of this gene in a comparison between primary and metastatic tumor cells in two-dimensional and three-dimensional culture conditions. In this study, after creating a mouse model of breast cancer using 4T1 cell line, primary breast cancer cells and brain and lung metastatic tumor cells were isolated and propagated in two-dimensional and three dimensional culture medium. Using real-time PCR reaction, analysis of nucleostemin gene expression was done comparetively between these two culture media. The findings of this experiment showed that the expression of the nucleostemin gene in the metastasis cycle in a two-dimensional culture medium is increased by 6 and 23 times, respectivly, in lung and brain metastatic tumor tissue compared to primary tumor cells. In the three-dimensional culture medium, which was done to enrich cancer stem cells, the expression level of nucleostemin gene in primary tumor cells and brain and lung metastatic cells compared to the two-dimensional culture medium in all three cell groups showed a significant decrease in expression. These findings provide information about nucleostemin gene expression in breast cancer metastatic cascade and 3D culture environment, which deserves further discussion. In this regard, analyzing the molecular properties of metastatic tumor cells, can be used to design targeted treatment strategies in the fight against breast cancer metastasis.

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