Mouse model of cachexia
Six-week-old male BALB/C mice (obtained from the Model Animal Research
Center of Nanjing University) were maintained under specific
pathogen-free (SPF) conditions and allowed to acclimate for one week.
Mice were housed in individually ventilated cages (IVCs) with free
access to drinking water and a basal diet under controlled conditions of
humidity, light (12 h light/12 h dark cycle), and temperature.
To induce the cachexia mouse model, 100 μl C26 murine adenocarcinoma
cells were subcutaneously inoculated into the right flanks of BALB/C
mice (5×105 cells/mouse). When the mice lost 20% of
their body weight or their tumor volumes reached 1500
mm3, the mice were sacrificed. Tumors, muscles, and
other organs, such as the liver and hypothalamus, were rapidly
dissected, frozen in liquid nitrogen and stored at -80°C until further
analyses or fixed with 10% formaldehyde for immunohistological
staining. All experiments involving animals were conducted according to
protocols approved by the Institutional Animal Care and Use Committee of
Nanjing University, in accordance with the guidelines for animal care
and use of Jiangsu Province.