Agilent LncRNA and mRNA array data for xenograft tumor of TE-2 cells in the nude mice

Although dysregulation of histamine and its receptor 4 (H4R) were widely and frequently appears in digestive carcinomas and was correlated with their malignancy and tumor proliferation, its existence and function in the esophagus and esophageal squamous cell carcinoma (ESCC) were still known. In our present study, we explored the expression and function of H4R in ESCC samples and cell lines. The results showed that H4R was overexpressed in poor differentiated ESCC samples and cell lines and correlated with median survival. H4R activation significantly not only blocked cell proliferation, cell cycle and invasion, but also induced the TE-2 xenograft inhibition and mice survival rate promotion. Mechanistic research demonstrated both metabolism (ACSS2) and non-metabolism (MAPK) related pathways were involved in H4R function, and H4R activation suppressed TGF-β1 via both above signaling. These findings firstly confirmed H4R overexpression in the esophagus cancer and its anti-tumor effects in the ESCC proliferation and invasion, which suggested that H4R was a potential target in therapy for ESCC. Agilent arrays were performed according to the manufacturer's directions on RNA extracted from xenograft tumors of TE-2. All animal procedures were performed following the National Animal Experimentation guidelines and were approved by the ethics committee of the Kunming General Hospital of Chengdu Military Region. All nu/nu mice were bred in aseptic conditions at a constant humidity and temperature with standard 12 h light-dark cycles and free access to drinking water and standard chow. TE-2 cells (1 × 107 in 100 mL of PBS) were suspended in a volume of 0.2 ml serum-free medium and injected subcutaneously into the hind flanks of the animals. At the end of the treatment, the nude rats were sacrificed and xenograft tumors were harvested for tumor volumes calculation (0.5 × length (mm) × width2 (mm2) as well as other experiments. When the subcutaneous tumors reached 20 mm3 (about 3 w), The mice were randomly assigned to one of the following four groups: (1) Vel group: nude mice received I.V. injections of vehicle (0.9% NaCl) once a day for 4 w; (2) 4MH group: nude mice received I.V. injections of 100 μM/kg H4R agonist 4MH once a day for 4 w; (3) JNJ group: nude mice received I.V. injections of 100 μM/kg H4R antagonist JNJ once a day for 4 w. Tissue samples and cells were put into liquid nitrogen for 10 min and then into an ultra freezer at 80°C. Total RNA was extracted using the GeneJET RNA purification Kit according to the manufacturer’s instructions (Thermo Scientific). RNA quantity and quality were measured by NanoDrop ND-1000. RNA integrity was assessed by standard denaturing agarose gel electrophoresis. Arraystar Mouse mRNA and LncRNA Microarray V3.0 is applied for the global profiling of mouse LncRNAs and protein-coding transcripts. About 35,923 LncRNAs and 24,881 coding transcripts can be detected by our third-generation mRNA and LncRNA microarray. Differentially expressed genes (DEGs) of 4MH group (between 100 μM/kg histamine H4 receptor agonist 4-methylhistamine treatment and control vehicle (0.9% NaCl) treatment) and JNJ group (between 100 μM/kg histamine H4 receptor antagonist JNJ7777120 and control vehicle (0.9% NaCl) treatment) were identified through the random variance model. A p-value was calculated using the paired t-test. The threshold set for up- and down-regulated genes was a fold change ≥ 2.0. The GO and pathway analysis were performed and the pain related GO terms were picked up for subsequent examination.