Rapamycin (Sirolimus)

Injection i.p.

U87-MG, T98G, and U373-MG

A Phase II study of Rapamycin to erase angiofibromas in patients with Tuberous Sclerosis Complex (TSC) is currently ongoing.

Rapamycin (Sirolimus, AY-22989, WY-090217) is a specific mTOR inhibitor with IC50 of ca.0.1 nM.

Dissolved in solvent solution (0.2% carboxymethylcellulose and 0.25% Tween-80 in sterile H2O)

Rapamycin inhibits endogenous mTOR activity in HEK293 cells with IC50 of ca.0.1 nM, more potently than iRap and AP21967 with IC50 of ca.5 nM and ca.10 nM, respectively. [1] In Saccharomyces cerevisiae, Rapamycin treatment induces a severe G1/S cell cycle arrest and inhibition of translation initiation to levels below 20% of control. [2] Rapamycin significantly inhibits the cell viability of T98G and U87-MG in a dose-dependent manner with IC50 of 2 nM and 1 uM, respectively, while displaying little activity against U373-MG cells with IC50 of >25 uM despite the similar extent of the inhibition of mTOR signaling. Rapamycin (100 nM) induces G1 arrest and autophagy but not apoptosis in Rapamycin-sensitive U87-MG and T98G cells by inhibiting the function of mTOR. [3]

72 hours

Cells are exposed to various concentrations of Rapamycin for 72 hours. For the assessment of cell viability, cells are collected by trypsinization, stained with trypan blue, and the viable cells in each well are counted. For the determination of cell cycle, cells are trypsinized, fixed with 70% ethanol, and stained with propidium iodide using a flow cytometry reagent set. Samples are analyzed for DNA content using a FACScan flow cytometer and CellQuest software. For apoptosis detection, cells are stained with the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) technique using an ApopTag apoptosis detection kit. To detect the development of acidic vesicular organelles (AVO), cells are stained with acridine orange (1 g/mL) for 15 minutes, and examined under a fluorescence microscope. To quantify the development of AVOs, cells are stained with acridine orange (1 g/mL) for 15 minutes, removed from the plate with trypsin-EDTA, and analyzed using the FACScan flow cytometer and CellQuest software. To analyze the autophagic process, cells are incubated for 10 minutes with 0.05 mM monodansylcadaverine at 37 C and are then observed under a fluorescence microscope.

Rapamycin (Sirolimus) Chemical Structure

Data from [J Lipid Res , 2011, 52, 1617-1625], Rapamycin (Sirolimus)purchased from Selleck, Rapamycin (RPM) inhibits OA-induced lipogenesis. Male SD rats were divided into three groups, and then treated as follows for seven days: group I (control, Ctrl), normal diet + vehicle, group II (OA), 1% OA diet + vehicle, group III (OA + RPM), 1% OA diet + RPM. (A) Hepatic TG concentrations were determined using the serum triglyceride determination kit. (B) Fixed liver sections were subjected to HE and Oil Red O staining as well as immunohistochemistry assays with SREBP-1 antibody. Representative microphotographs of liver specimens from all groups of rats are shown. (C) The levels of mRNA of the indicated genes involved in lipogenesis were determined by real-time PCR. Each bar represents the mean SE of the results obtained from six rats. * P, 0.05, ** P, 0.01, *** P, 0.001.

2 years -20CPowder, 2 weeks4Cin DMSO, 2 months-80Cin DMSO