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Taxanes are the first line of treatment for a wide range of cancers, including breast, lung and prostate cancer. Taxanes are very effective chemotherapy drugs, but we don’t fully understand how they work.
Taxanes are known to bind to microtubules, hollow structures that are a core component of cytoskeleton, or the cell’s internal skeleton.
Microtubules support cells to maintain their shape, move, transport materials internally and separate chromosomes during cell division.
It can be visualised using electron microscopy that on binding, taxanes change the structure of microtubules on the nanoscale.
Researchers found that this change causes a signalling protein called GEF-H1 to detach from the microtubules and activate the RhoA signalling pathway.
This signalling pathway reorganises actin, another part of the cytoskeleton that acts like the cell’s muscles, causing the cell to contract and change shape.
Insights into the mode of action of taxanes may lead to the design of better cancer therapies.
Cells were imaged on a custom Yokogawa spinning disc with an Eclipse Ti-E body (Nikon) with perfect focus, a MS-2000-XYZ stage with Piexo Top Plate (ASI), a CSU-X1-A1 spinning disc unit (Yokogawa) and STXG-PLAMX-SETZ21L temperature and CO2 control (TokaiHit). An S Fluor ×100 oil objective (Nikon) was used to acquire images. Voltran Stradus 488-nm (Vortran) and Coherent OBIS 561-nm (Coherent) lasers were used to excite the sample, and ET525/50m (GFP) and ET630/75m (mCherry) filters were used to filter the signal. A prime BSI sCMOS camera (Teledyne Photometrics) was used to take images, and MetaMorph 7.10 (Molecular Devices) software was used to control the microscope.
Microscope incubation chambers were preheated to 37 °C with 5% CO2. One image was taken every 30 s (GFP-GEF-H1 imaging) or 1 min (F-actin imaging); the first 10 min acquired served as a baseline, after which either 10 µM paclitaxel (PTX) or discodermolide (DDM) was applied to cells and cells were imaged for another 10–15 min or 1 h.
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