Fig. 7
Knockdown of Hotair in MBT-2 bladder tumor-bearing mice attenuates cisplatin-induced cancer cachexia. a Treatment schedule. Groups of male C3H/HeN mice were subcutaneously inoculated with 3 × 106 of MBT-2, HOTAIR-knockdown MBT-2 (MBT-2/CSi-Hotair-F4 and -F6), or GFP-knockdown MBT-2 (MBT-2/CSi-GFP) cells at day 0. Another group of mice that received vehicle treatment served as the healthy control. Except for healthy control mice, all mice were treated intraperitoneally with cisplatin (5 mg/kg/day) at days 17, 19, 21, 23, 25 and 27. The mice were sacrificed at day 30. b Body weight (after subtraction of the estimated tumor weight, which was estimated as 1 mm3 tumor volume equal to 1 mg) was expressed as a percentage of the body weight at day 0. c Body weight at day 29 (after subtraction of the absolute tumor weight) was expressed as a percentage of the body weight at day 0. d Tumor volumes were measured every two days. Values shown are mean ± SD (n = 4–6) (b–d). e Serum levels of IL-6 was quantified by ELISA (n = 5–9). f Calf circumference was measured at day 30 before sacrifice (n = 2–6). g Cross-sectional areas of H&E-stained gastrocnemius muscle tissues (upper) and quantitation using the ImageJ software (lower) (n = 3–6). Scale bar = 50 μm. h Immunohistochemistry (upper) and quantitation (lower) of MuRF-1 in muscle tissue sections (n = 2–6). Scale bar = 200 μm