In the present study, we observed that mTOR and P70S6K expression were examined in gastric carcinoma, adjacent non-tumorous mucosaand adenoma, and compared with the clinicopathological
parameters of tumors to explore the clinicopathological significance and molecular role of the mTOR signal pathway in the stepwise development of gastric carcinomas. Materials and methods Subjects Gastric carcinomas (n = 421) were collected from the surgical resection, adenoma (n = 45) from endoscopic biopsy or polypectomy, and gastritis selleck chemicals (n = 49) from the endoscopic biopsy in Shengjing Hospital of China Medical University and the First Affiliated Hospital of China Medical University between 1993 and 2006. All carcinomas were adenocarcinomas and the adenoma group was free from non-neoplastic polyp types, leiomyomas and benign GIST’s. The patients with gastric carcinoma were 293 men and 126 women (29~91 years, mean = 65.4 years). Among them, 156 cases have carcinomas accompanied with lymph node metastasis. None of the patients underwent chemotherapy or radiotherapy before surgery. They all provided consent for use of tumour tissue for clinical research and our University
Ethical Committee approved the research protocol. We followed up all patients by consulting their case documents or through telephone. Pathology All tissues were fixed in 4% neutralised formaldehyde, embedded in paraffin and incised into 4 mm sections. These sections Fedratinib order were Selleck AZD8186 stained by haematoxylin-and-eosin (HE) to confirm their histological diagnosis and other
microscopic characteristics. The staging for each gastric carcinoma was evaluated according to the Union Internationale Contre le Cancer (UICC) system for the extent of tumour spread . Histological architecture of gastric carcinoma was expressed in terms of Lauren’s classification [13, 14]. Furthermore, tumour size, depth of invasion, lymphatic and venous invasion were determined. Tissue microarray U0126 (TMA) Prior to TMA construction, all tissue slides were histopathologically re-evaluated by one pathologist and. Two 2.0-mm tissue cores were taken from representative areas of gastric samples using a manual arraying device (MTA-1; Beecher Inc., Sun Prairie, WI, USA) and mounted in a new recipient block. Four-μm-thick sections were consecutively incised from the recipient block and transferred to poly-lysine-coated glass slides. HE staining was performed on TMA for confirmation of tumor tissue. Immunohistochemistry For the immunohistochemical procedure, 4-μm-thick sections were deparaffinized with xylene and rehydrated through an alcohol gradient. The sections were quenched with 3% hydrogen peroxide in absolute methanol for 20 min to block endogenous peroxidase activity, and heated in a microwave for 15 min in citrate buffer (0.01 mol/L, pH 6.0) to retrieve the antigen.