Roles of Endocytic Processes and Early Endosomes on Focal Adhesion Dynamics in MDA-MB-231 Cells.

Bandar Fahad Alharbi,Dhurgham Al-Fahad,Philip Richard Dash

doi:10.52547/rbmb.10.2.145

Bandar Fahad Alharbi, Dhurgham Al-Fahad + Show 1 more

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https://doi.org/10.52547/rbmb.10.2.145

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Abstract

Focal adhesion (FA) play a critical role in many biological processes which include cell survival and cell migration. They serve as cellular anchor, allowing cells to stay attached to the extracellular matrix (ECM), and can also regulate cellular transduction. Previously, it has been suggested that vesicles such as endosomes could interact directly with FA or be implicated in their turnover. In this study, we investigated whether there is a relationship between FA and the early endocytic machinery in MDA-MB-231 cells. In this study, cell culture, transfection, time laps confocal microscopies, immunocytochemistry, western blotting, Cell fractionation and immunoprecipitation techniques were performed. Cells acutely treated with Dynasore, an inhibitor of dynamin, or with Pitstop 2, an inhibitor of clathryn-dependent endocytosis showed a reduction in the expression of early endosome biomarkers such as Rab5 and EEA1. Additionally, cells treated with these endocytic inhibitors exhibited an increase number and size of FA, as well as an increase FA turnover duration. This data was consistent with the reduction of the speed of cell migration. We demonstrated that Rab5- and EEA1-positive early endosomes were found to be colocalized with internalized FA. The present study suggests that there is a link between FA and early endosome markers, which indicates that the early endosomes may be involved in FA dynamics.

Highlights

The miscarriage is a common phenomenon that usually occurs in the first trimester of pregnancy between 15% -17% of pregnancies, but the true percentage is likely higher because many miscarriages occur early in the pregnancy without the women even realizing that they are pregnant [1]
It has been demonstrated that B19 infection is associated with elevated circulating levels of interferon and interleukins that are linked with several diseases [15]
Little attention has been given to evaluate the levels of interleukin 6 (IL-6), IL-8 and IFN-γ in aborted women infected (B19+/Abr+) and uninfected (B19-/Abr+) with B12

Summary

Introduction

The miscarriage is a common phenomenon that usually occurs in the first trimester of pregnancy between 15% -17% of pregnancies, but the true percentage is likely higher because many miscarriages occur early in the pregnancy without the women even realizing that they are pregnant [1]. RhD and Cytokine against B19 in Miscarriage remains inside the uterus. This condition is associated with vaginal bleeding of varying degrees of severity. It has been reported that’s susceptibility of the ABO blood type people to several viral infection. We evaluated interleukin 6 (IL-6), interleukin 8(IL-8), and interferon gamma (IFN-γ) levels in aborted women infected with parvovirus B19 (B19+/Abr+) and uninfected with B19(B19-/Abr+) in comparison with healthy women (B12-/Abr-) and susceptibility of their RhD blood type to contract B19. Our analyzed data showed that aborted women with RhD+ are more susceptible to contract s B19 than people with RhD- blood type. The susceptibility/protection of aborted women against B19 might be determined based on RhD blood type

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