Cho Nam Hoon, Hong Kyi Pyo, Hong Sung Hui, Kang Suki, Chung Kyung Young, Cho Sang Ho
Department of Pathology, Yonsei University College of Medicine, Seoul, Korea.
Oncogene. 2004 Jan 22;23(3):845-51. doi: 10.1038/sj.onc.1207140.
We aimed to clarify the prime role of recurrence in stage I lung cancer. To determine the expression profiles, quantitative RT-PCR and real-time PCR were performed subsequently to evaluate the validity of meaningful molecules identified by 0.12 K c-DNA array experiment surveys. In all, 10 lung cancer patients presenting with recurrence of stage IB were selected and compared with 10 stage IB lung cancer patients without recurrence since biopsied 3 years previously. On c-DNA microarray data analysis using pairs of recurred and the corresponding nonrecurred patients, the following genes were found to be upregulated in the recurred cases: matrix metalloproteinase (MMP)-10 in five cases, MMP-12 in two cases, MMP-11, MMP-14, MMP-15, fos, cyclin E2, E2F3, TGF-alpha in each one case. The most frequently upregulated genes in recurred lung cancers were MMP-10 (stromelysin-2) and MMP-12 (macrophage elastase). On transcriptional assay by quantitative RT-PCR and real-time RT-PCR analysis to validate those molecules, both transcripts of MMP-10 and MMP-12 were significantly more upregulated in recurred stage IB lung cancer than in the non-recurred stage IB lung cancer (P=0.004). Transcript levels were identical to c-DNA array data. The protein levels of these entities were also evaluated by immunohistochemistry of archival slides. By immunohistochemistry, MMP-10 monoclonal antibody showed more intense immunoreactivity in the recurred stage IB lung cancer than in the nonrecurred stage IB lung cancer (P=0.0313). Our approach revealed that MMP-10 plays an important role in the recurrence in stage IB lung cancer, irrespective of the histologic type.
我们旨在阐明复发在I期肺癌中的主要作用。为了确定表达谱，随后进行了定量逆转录聚合酶链反应（qRT-PCR）和实时聚合酶链反应（real-time PCR），以评估通过0.12K cDNA阵列实验检测鉴定出的有意义分子的有效性。总共选择了10例出现IB期复发的肺癌患者，并与10例自3年前活检后未复发的IB期肺癌患者进行比较。在使用复发患者与相应未复发患者对的cDNA微阵列数据分析中，发现以下基因在复发病例中上调：5例中的基质金属蛋白酶（MMP）-10、2例中的MMP-12、各1例中的MMP-11、MMP-14、MMP-15、fos、细胞周期蛋白E2、E2F3、转化生长因子-α（TGF-α）。复发肺癌中最常上调的基因是MMP-10（基质溶解素-2）和MMP-12（巨噬细胞弹性蛋白酶）。通过定量RT-PCR和实时RT-PCR分析进行转录测定以验证这些分子，MMP-10和MMP-12的转录本在复发的IB期肺癌中均比未复发的IB期肺癌上调更显著（P = 0.004）。转录水平与cDNA阵列数据一致。这些实体的蛋白质水平也通过存档玻片的免疫组织化学进行评估。通过免疫组织化学，MMP-10单克隆抗体在复发的IB期肺癌中显示出比未复发的IB期肺癌更强的免疫反应性（P = 0.0313）。我们的方法表明，无论组织学类型如何，MMP-10在IB期肺癌复发中起重要作用。
