Thanasai Jongkonnee, Limpaiboon Temduang, Jearanaikoon Patcharee, Bhudhisawasdi Vajarabhongsa, Khuntikeo Narong, Sripa Banchob, Miwa Masanao
Graduate School, Faculty of Medical Technology, Mahidol University, Bangkok 10700, Thailand.
World J Gastroenterol. 2006 Jul 21;12(27):4338-44. doi: 10.3748/wjg.v12.i27.4338.
To analyze the DNA copy number of target genes NF2, TIMP3, ST13, TOB2, BIK, and TP and the reference microsatellite markers D22S283, D22S423, and D22S274 mapped on 22q12-qter in liver fluke related cholangiocarcinoma (CCA) and define its correlation with clinical parameters.
Quantitative real time PCR (qPCR) was used for determining allelic imbalances in 65 liver fluke related CCA tissues. Statistical correlations between allelic imbalances and clinicopathological parameters, i.e. age, sex, tumor stage, histological type, blood vessel invasion, nerve invasion and lymphatic invasion were evaluated by means of the chi2 test. Cox regression analysis was used for determining patient's survival.
Amplifications of the TP (22q13.33), TOB2 (22q13.2-13.31), D22S283 (22q12.3), TIMP3 (22q12.3) and NF2 (22q12.2) were found in 35 (53.8%), 28 (43.1%), 27 (41.5%), 24 (36.9%), and 24 (36.9%), respectively. Losses at the D22S423 (22q13.1-13.2) and BIK (22q13.31) were detected in 26 (40%) and 23 (35.4%), respectively. Significant correlations were observed between lymphatic invasion and allelic losses of BIK (P = 0.025) and D22S283 (P = 0.041). Univariate and multivariate Cox regression analysis revealed D22S283 amplification as an independent predictor of good prognosis (P = 0.006, death hazard ratio = 0.411, 95% CI = 0.217-0.779) and blood vessel invasion as an independent poor prognostic factor (P = 0.042, death hazard ratio = 1.911, 95% CI = 1.022-3.571) in CCA patients.
This study provides evidence for the involvement of gene amplification and deletion on chromosome 22q in liver fluke related CCA. This is the first report of D22S283 amplification as an independent indicator of favorable prognosis in liver fluke related CCA.
分析肝吸虫相关性胆管癌（CCA）中靶基因NF2、TIMP3、ST13、TOB2、BIK和TP以及定位在22q12 - qter的参考微卫星标记D22S283、D22S423和D22S274的DNA拷贝数，并确定其与临床参数的相关性。
采用定量实时PCR（qPCR）检测65例肝吸虫相关性CCA组织中的等位基因失衡情况。通过卡方检验评估等位基因失衡与临床病理参数（即年龄、性别、肿瘤分期、组织学类型、血管侵犯、神经侵犯和淋巴侵犯）之间的统计学相关性。采用Cox回归分析确定患者的生存率。
分别在35例（53.8%）、28例（43.1%）、27例（41.5%）、24例（36.9%）和24例（36.9%）中发现TP（22q13.33）、TOB2（22q13.2 - 13.31）、D22S283（22q12.3）、TIMP3（22q12.3）和NF2（22q12.2）的扩增。在26例（40%）和23例（35.4%）中分别检测到D22S423（22q13.1 - 13.2）和BIK（22q13.31）的缺失。观察到淋巴侵犯与BIK（P = 0.025）和D22S283（P = 0.041）的等位基因缺失之间存在显著相关性。单因素和多因素Cox回归分析显示，D22S283扩增是CCA患者良好预后的独立预测因子（P = 0.006，死亡风险比 = 0.411，95%可信区间 = 0.217 - 0.779），血管侵犯是独立的不良预后因素（P = 0.042，死亡风险比 = 1.911，95%可信区间 = 1.022 - 3.571）。
本研究为22号染色体上基因扩增和缺失参与肝吸虫相关性CCA提供了证据。这是首次报道D22S283扩增作为肝吸虫相关性CCA良好预后的独立指标。
