Invadopodia Formation is a Critical Step in Cancer Cell Invasion: The Effect of Passage Number on Invadopodia Formation in MDA-MB-231 Breast Cancer Cell Line

Hamad Ali Hamad,Nur Fariesha Md Hashim,Yoke Kqueen Cheah

doi:10.28916/lsmb.2.3.2018.20

Hamad Ali Hamad, Nur Fariesha Md Hashim + Show 1 more

Open Access

https://doi.org/10.28916/lsmb.2.3.2018.20

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Abstract

High invasive cancer cells are thought to recruit specialised actin-rich protrusions for invasion in metastasis process. These protrusions are termed invadopodia. To study invadopodia formation, one of the first challenges faced by researchers has been to optimise the cell line passage number in order to be used for the invadopodia assay. Therefore, this study aims to investigate the effects of the passage number on invadopodia formation in MDA-MB-231 breast cancer cell line. Invadopodia assay was used to achieve the aim of the study. The results provided evidence that invadopodia formation is affected by the high passage number. The cells were also tested with dimethyloxalylglycine (DMOG) a hypoxic mimicking agent which is known to be an invadopodia inducer, the results showed that the cells in low passage number (P7) treated with DMOG increase the cells forming invadopodia, while the cells with high passage number (P35) showed that DMOG fails to stimulate the cells to form invadopodia. Furthermore, the cells with high passage number after passage 15 are starting to lose the ability to degrade the gelatin. In conclusion, this study suggests that only cells with a low passage number, less than passage 15 should be used in the study of invadopodia formation to obtain the results in the search for molecular targets and signaling at invadopodia.

Highlights

Breast carcinoma cells need to invade the surrounding extracellular matrix (ECM) to achieve metastasis successfully
The cells were tested with dimethyloxalylglycine (DMOG) a hypoxic mimicking agent which is known to be an invadopodia inducer, the results showed that the cells in low passage number (P7) treated with DMOG increase the cells forming invadopodia, while the cells with high passage number (P35) showed that DMOG fails to stimulate the cells to form invadopodia
The present study aims to use invadopodia assay to investigate the effects of passage number on invadopodia formation in MDA-MB-231 cell line

Summary

Introduction

Breast carcinoma cells need to invade the surrounding extracellular matrix (ECM) to achieve metastasis successfully. The majority of cancer mortalities are primarily due to metastasis (Xie et al, 2017), investigation of the mechanism of cancer cells to invade the ECM during metastatic cascade is critical in cancer study (Bowden et al, 1999). Actin-rich subcellular protrusions have been shown to be critical for ECM degradation. These protrusions are termed invadopodia and are formed in the ventral of highly invasive cancer cells. In a study reported by Hashim et al, (2013), the invasiveness of MDA-MB-231 breast cancer cells increases due to increased expression of the hypoxia-inducible factor-1α (HIF1α) under hypoxic environment.

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