Identification of genes differentially expressed in a newly isolated human metastasizing esophageal cancer cell line, T.Tn‐AT1, by cDNA microarray

Abstract

sophageal squamous cell carcinoma (SCC) is one of the most aggressive cancers. Esophageal SCC is relatively common in countries in Eastern Asia, such as China and Japan, and is characterized by poor prognosis and rapid clinical progression, with a high frequency of lymph node metastasis and recurrence. The 5-year survival rate of patients with esophageal SCC showing submucosal invasion is low (40-75%) 1-5) compared with that for patients with colon cancer (over 80%). 4,5) In addition, lymph node metastasis is commonly found in esophageal SCC, even when the tumor invades only the submucosa. Lymph node metastasis is the main cause of the poor prognosis for the patients with esophageal SCC.Consequently, the identification of the genes associated with metastasis of esophageal SCC is very important. Microarray analysis has been used to investigate the gene expression profiles of esophageal SCC tissues and esophageal cancer cell lines with various characteristics.6-10) When clinical materials are used for microarray analysis, it might be necessary to look at the overall expression profiles of genes (if possible) by clustering analysis in the different pathological stages, such as dysplasia, carcinoma in situ, and invasive cancer with or without metastasis. However, to isolate gene(s) related to metastasis, it would be convenient to use cell lines with different metastatic potentials derived from the same parental cells, and a reliable model system is therefore required for examining the metastatic potential of cancer cells.In our previous experiments, we established in vitro and in vivo model systems for studying invasion and metastasis of esophageal SCC cells. 11,12) We first clarified in detail the molecular and genetic characteristics of a human non-metastasizing esophageal SCC cell line, T.Tn, 12) and then developed an orthotopic inoculation model for esophageal cancer cells in nude mice.11) In the present study, we isolated a metastasizing subclone from the parental non-metastasizing T.Tn cell line by in vitro selection and by the use of a nude mouse orthotopic inoculation model. Then, we compared the expression profiles of 9206 genes in the parental T.Tn cells and the metastasizing subclone by cDNA microarray analysis, and identified several genes differentially expressed in the metastasizing subclone. Materials and MethodsCell line. A human esophageal SCC cell line, T.Tn 13) was obtained from JCRB (Japanese Collection of Research Bioresources, Osaka). T.Tn cells were grown in 1:1 mixture of Dulbecco's modified Eagle's medium (Nissui, Tokyo) and F-12 (Life Technologies, Inc., Gaithersburg, MD) supplemented with 10% fetal calf serum (FCS; Sigma, St. Louis, MO), 100 µg/ml streptomycin, 100 units/ml penicillin (Life Technologies, Inc.), and 0.25 µg/ml amphotericin B (Life Technologies, Inc.) in a humidified atmosphere of 95% air and 5% CO 2 at 37°C. HT1080 cells are derived from a human fibrosarcoma cell line known to secrete a large amount of several matrix-degrading enzymes (purchased from Dai-Nippon Seiyaku, Osa...

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F-Box Only Protein 31 (FBXO31) Negatively Regulates p38 Mitogen-activated Protein Kinase (MAPK) Signaling by Mediating Lysine 48-linked Ubiquitination and Degradation of Mitogen-activated Protein Kinase Kinase 6 (MKK6)

“…The ESCC 2 cell line T.Tn was cultured in DMEM/F12 (Invitrogen) supplemented with 10% FBS (20). HeLa cells, HEK293 cells, and the packaging cells 293 Phoenix and 293T were cultured in DMEM containing 10% FBS.…”

Id1-Induced IGF-II and Its Autocrine/Endocrine Promotion of Esophageal Cancer Progression and Chemoresistance—Implications for IGF-II and IGF-IR–Targeted Therapy

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