Upregulation of topoisomerase IIalpha expression in advanced gallbladder carcinoma: a potential chemotherapeutic target.
OBJECTIVE
The lack of treatment options other than surgical resection results in unfavourable prognosis of advanced gallbladder carcinoma. The aim of this study was to identify cancer-specific cellular targets that would form the basis for some therapeutic approaches for this disease.


METHODS
Twelve advanced gallbladder carcinoma tissue samples and three samples of normal gallbladder epithelium were screened to identify differentially expressed genes by DNA microarray analysis. The results obtained were validated in an independent sample set by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR). Among the genes picked-up, one molecule, topoisomerase IIalpha (TOPO IIalpha), was further assessed immunohistochemically as a potential chemotherapeutic target, and the growth inhibitory effects of etoposide, doxorubicin and idarubicin, representative TOPO IIalpha inhibitors, on two different gallbladder carcinoma cell lines were compared with that of gemcitabine and 5-fulorouracil.


RESULTS
Five upregulated genes were identified: four cell cycle-related genes (TOPO IIalpha, cyclin B2, CDC28 protein kinase regulatory subunit 2, ubiquitin-conjugating enzyme E2C) and a metabolism-related gene (gamma-glutamyl hydrolase). Immunohistochemically, TOPO IIalpha was expressed in gallbladder cancer cells, and 16 of 35 cases (46%) had strong TOPO IIalpha expression defined as having a labeling index of >50%. In in vitro growth inhibition assay, etoposide, as well as doxorubicin and idarubicin, was the most effective for OCUG-1 cells that had strong TOPO IIalpha expression, while gemicitabine was the most effective for NOZ cells with weak TOPO IIalpha expression. Etoposide induced apoptosis of OCUG-1 cells.


CONCLUSIONS
TOPO IIalpha might be an effective chemotherapeutic target in advanced gallbladder carcinoma, especially when it is expressed strongly.
Washiro
Mitsutsune
M
Department of General Surgery, Graduate School of Medicine, Chiba University, 1-8-1 Inohana, Chuoh-ku, Chiba, 260-8670, Japan.
Ohtsuka
Masayuki
M
Kimura
Fumio
F
Shimizu
Hiroaki
H
Yoshidome
Hiroyuki
H
Sugimoto
Takashi
T
Seki
Naohiko
N
Miyazaki
Masaru
M
D000368:Aged; D000951:Antigens, Neoplasm / Q000096:biosynthesis* / Q000235:genetics; D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology; D014408:Biomarkers, Tumor / Q000032:analysis; D015153:Blotting, Western; D002277:Carcinoma / Q000235:genetics* / Q000378:metabolism*; D049109:Cell Proliferation / Q000187:drug effects; D004250:DNA Topoisomerases, Type II / Q000096:biosynthesis* / Q000235:genetics; D004268:DNA-Binding Proteins / Q000096:biosynthesis* / Q000235:genetics; D003841:Deoxycytidine / Q000031:analogs & derivatives / Q000494:pharmacology; D004317:Doxorubicin / Q000494:pharmacology; D005047:Etoposide / Q000494:pharmacology; D005260:Female; D005706:Gallbladder Neoplasms / Q000235:genetics* / Q000378:metabolism*; D015870:Gene Expression; D020869:Gene Expression Profiling; D006801:Humans; D015255:Idarubicin / Q000494:pharmacology; D007150:Immunohistochemistry; D008297:Male; D020411:Oligonucleotide Array Sequence Analysis; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015854:Up-Regulation; D000093542:Gemcitabine
D000951:Antigens, Neoplasm; D000970:Antineoplastic Agents; D014408:Biomarkers, Tumor; D004268:DNA-Binding Proteins; D012333:RNA, Messenger; D003841:Deoxycytidine; D005047:Etoposide; D004317:Doxorubicin; D004250:DNA Topoisomerases, Type II; D015255:Idarubicin; D000093542:Gemcitabine