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Fig. 6 | Journal of Biomedical Science

Fig. 6

From: Interruption of the long non-coding RNA HOTAIR signaling axis ameliorates chemotherapy-induced cachexia in bladder cancer

Fig. 6

The CM of HOTAIR-overexpressing bladder cancer cells, in particular, in the presence of cisplatin promotes muscle atrophy. Murine C2C12 myoblasts were induced to differentiate into myotubes by culturing the cells in DMEM supplemented with 2% horse serum for 4 days. The CM containing 10% FBS collected from cancer cells was diluted 1:1 with fresh DMEM containing 2% horse serum to treat myotubes for 48 h. a, b C2C12 myotubes were treated with 100 ng/ml of mouse TNF-α (mTNF-α), human TNF-α (hTNF-α), or human IL-6 (hIL-6) in DMEM containing 5% FBS and 1% horse serum for 48 h. c, d The CM collected from J82/HOTAIR, J82/GFP, or parental (mock) J82 cells in the absence or presence of cisplatin (2 μg/ml) for 48 h was used to treat C2C12 myotubes. e. f The CM collected from MBT-2/CSi-Hotair-F4, MBT-2/CSi-Hotair-F6, MBT-CSi-GFP, or parental (mock) MBT-2 cells in the absence or presence of cisplatin (2 μg/ml) for 48 h was used to treat C2C12 myotubes for 48 h. A total of 105 myotubes (15 myotubes/filed) within each section were measured in randomly selected seven different view fields using the ImageJ software to calculate the average diameters. C2C12 representative images (a, c, e) and average diameter of myotubes (b, d, f). Scale bar = 100 μM. g, h Detection of muscle-specific E3 ubiquitin ligases MuRF-1, UBR2, and atrogin-1 in C2C12 myotubes after treatment with the CM of MBT-2 cells or their derivatives by immunoblotting. Expression of β-actin served as the loading control. Values shown below the blots are ratios between the intensity of the bands corresponding to the indicated proteins and those corresponding to β-actin analyzed by densitometry

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