Iron overload is related to muscle wasting in patients with cachexia of gastric cancer: using quantitative proteome anal
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ORIGINAL PAPER
Iron overload is related to muscle wasting in patients with cachexia of gastric cancer: using quantitative proteome analysis Da Zhou1 · Ying Zhang2 · Gulsudum Mamtawla1 · Songlin Wan1 · Xuejin Gao1 · Li Zhang1 · Guoli Li1 · Xinying Wang1 Received: 16 August 2020 / Accepted: 30 October 2020 © Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract The incidence of gastric cancer cachexia is high and the clinical management is poor, so the study aimed to clarify the mechanism of muscle wasting to better screen patients with gastric cancer cachexia. Gastric cancer patients undergoing radical gastrectomy were divided into cachexia with sarcopenia (CS, n = 13) and normal (N, n = 10) two groups. The possible mechanism of skeletal muscle reduction was explored through Tandem Mass Tag (TMT) technique, Perls staining, Western blot analysis and measurement of oxidative stress indicators. The preoperative weight, weight loss, body mass index, calorie intake and skeletal muscle index values of the CS group were significantly lower than those of the N group (P 5% or body mass index (BMI) 2% or sarcopenia and weight loss > 2% were defined as cachexia [5]. Patients with gastric cancer of any gender between 40 and 70 years old were divided into two groups: cachexia with sarcopenia (CS) or normal (N) group according to the above definition. All patients were volunteer to participate in the study. The patients have given their written informed consent, and the study was approved by the Ethics Committee of Jinling Hospital (approval no. 2017NZKY-003-01). The inclusion and exclusion criteria of patients refer to our previous studies
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Medical Oncology
(2020) 37:113
[11], and patients with a history of using iron agents or iron chelators, long-term or heavy alcohol drinking (drinking for more than 5 years, with an average daily consumption of more than 30 g) were also excluded. Finally, the abdominal muscles of 23 patients were biopsied, and we collected their demographic and clinical data, including age, gender, height, weight and weight loss (including causes), BMI, history of smoking and alcohol drinking, comorbidities, SMI, calorie intake, serum nutritional indicators (insulin-like growth factor-1, albumin, prealbumin, transferrin, and retinol-conjugated protein) and serum ferritin levels.
Muscles biopsies Surgical muscle biopsies were taken immediately after opening the abdominal cavity during radical gastrectomy and about 500 mg of muscle tissue was extracted using a scalpel. Biopsies were immediately frozen in liquid nitrogen and stored at − 80 °C.
Sample preparation One milliliter of lysis buffer [7 M urea, 4% SDS, 1 × Protease Inhibitor Cocktail (Roche Ltd. Basel, Switzerland)] was added to samples, followed by sonication on ice and centrifugation at 13,000 rpm for 10 min at 4 ℃. The supernatant was transferred to a fresh tube.
Protein digestion and TMT labeling and high pH reverse phase separation Determine the protein concentration of the supernatant using the BCA protein ass
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