Mori Kazuhiko, Aoyagi Kazuhiko, Ueda Tetsuya, Danjoh Inaho, Tsubosa Yasuhiro, Yanagihara Kazuyoshi, Matsuno Yoshihiro, Sasako Mitsuru, Sakamoto Hiromi, Mafune Ken ichi, Kaminishi Michio, Yoshida Teruhiko, Terada Masaaki, Sasaki Hiroki
Genetics Division, National Cancer Center Research Institute, Tokyo 104-0045, Japan.
Biochem Biophys Res Commun. 2004 Jan 23;313(4):931-7. doi: 10.1016/j.bbrc.2003.12.025.
Peritoneal wash cytology plays a pivotal role in the decision for gastric cancer treatment because advanced gastric cancer often turns out incurable with peritoneal metastasis. Molecular detection of minimal cancer cells from peritoneal washings may overcome the sensitivity boundary of conventional cytology and contribute to the prediction of the disease outcome. To select marker candidates out of ten thousands of genes, we performed microarray analyses in 12 gastric cell lines and 8 peritoneal washings of early stage cases. With 40 candidates selected by the above expression profiling, RT-PCR in 16 representative peritoneal wash samples was performed to identify genes specific to cytology positive samples. The finally selected five genes, CK20, FABP1, MUC2, TFF1, and TFF2, were then evaluated for their utility as a marker for minimal residual disease in 99 peritoneal wash samples. Nested RT-PCR using the five genes showed positive results highly specific to incurable cases (91-100%). With a high specificity, the combination of these five genes succeeded in identifying 6 out of 20 (30%) additional patients with all types of early recurrence that could not be predicted by the conventional method. The six newly identified recurrences included four non-peritoneal ones, showing that RT-PCR using the five genes without a real-time quantitative PCR technique contributes to the detection of minimal residual disease.
腹膜冲洗细胞学检查在胃癌治疗决策中起着关键作用，因为晚期胃癌往往因腹膜转移而无法治愈。从腹膜冲洗液中分子检测微小癌细胞可能会突破传统细胞学的敏感性界限，并有助于预测疾病预后。为了从数以万计的基因中筛选出候选标志物，我们对12种胃癌细胞系和8例早期病例的腹膜冲洗液进行了微阵列分析。通过上述表达谱分析筛选出40个候选基因后，对16个代表性腹膜冲洗样本进行了逆转录聚合酶链反应（RT-PCR），以鉴定细胞学阳性样本特有的基因。然后，对最终筛选出的5个基因，即细胞角蛋白20（CK20）、脂肪酸结合蛋白1（FABP1）、黏蛋白2（MUC2）、三叶因子1（TFF1）和三叶因子2（TFF2），在99个腹膜冲洗样本中评估其作为微小残留病标志物的效用。使用这5个基因进行巢式RT-PCR显示，结果对不可治愈病例具有高度特异性（91%-100%）。这5个基因的组合以高特异性成功识别出20例（30%）所有类型早期复发患者中的6例，这些患者用传统方法无法预测。新发现的6例复发包括4例非腹膜复发，表明在不使用实时定量PCR技术的情况下，使用这5个基因进行RT-PCR有助于检测微小残留病。
