CD44-mediated binding and internalization of high and low molecular weight hyaluronic acid in malignant mesothelioma cells

CD44-mediated binding and internalization of high and low molecular weight hyaluronic acid in malignant mesothelioma cells

Cell receptors

Hyaluronic acid -- Physiological effect

Ligands (Biochemistry)

Mesothelioma

Poston, Stacie Jean

2008

Seischab, Lori:

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Low molecular weight hyaluronan (HA) is found in high concentrations in the serum of patients with malignant mesothelioma. It has also been shown previously to induce migration of certain cells, and induce specific signaling pathways. CD44 expressed on the cell surface serves as the primary receptor for HA. A modified Boyden chamber assay consisting of HA-embedded filters was used to study the role of HA size in migration of malignant mesothelioma cells (CRL-5946 and CRL-5919). Cell migration over high and low molecular weight HA was similar. Control conditions lead to mixed results where cells incubated with antibodies (Ab) against CD44 showed less migration (15.1% average decrease) in some cases. While in other repetitions, cells treated with anti-CD44 Abs migrated more readily than cells with no Ab present. Anti-CD44 Ab may lead to enhanced binding of HA through CD44 clustering. Fluorescein attached to HA via a biotin-avidin link was used to measure the differential binding of high and low molecular weight HA to malignant mesothelioma cells. Fluorescence was measured using a microtiter plate reader. Fluorescence levels varied on average by less than 3.0% between treated and control cells. A similar assay with fluorescein-labeled HA was used to determine internalization over 3 hr and 24 hr of low and high molecular weight HA. Fluorescence levels varied on average by less than 0.1% between treated and control cells. It is possible that using assays based on microtiter plate readers may be ineffective in studying binding and internalization of HA by malignant mesothelioma cells due to the scatter produced in control systems.

2015-06-08

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