Toiyama Yuji, Mizoguchi Akira, Kimura Kazushi, Hiro Junichirou, Inoue Yasuhiro, Tutumi Tomonari, Miki Chikao, Kusunoki Masato
Department of Gastrointestinal and Pediatric Surgery, Division of Reparative Medicine, Institute of Life Sciences, Mie University Graduate School of Medicine, 2-174 Edobashi, Tsu, Mie 514-8507, Japan.
World J Gastroenterol. 2007 May 21;13(19):2717-21. doi: 10.3748/wjg.v13.i19.2717.
To investigate the expression patterns of TTYH2 in the human colon cancer and colon cancer cell lines and to evaluate the inhibitory effect of small interfering RNA (siRNA) on the expression of TTYH2 in colon cancer cell lines.
We investigated the expression patterns of TTYH2 in colon cancer, adjacent non-tumorous colon mucosa, and cancer cell lines (DLD-1, caco-2, and Lovo) by RT-PCR. Furthermore, a siRNA plasmid expression vector against TTYH2 was constructed and transfected into DLD-1 and Caco-2 with Lipofectamine 2000. The down regulation of TTYH2 expression was detected by RT-PCR and the role of siRNA in inducing cell proliferation and cell aggregation was evaluated by MTT and aggregation assay.
TTYH2 gene expression in colon cancer tissue was significantly up-regulated compared with normal colonic mucosa (1.23 +/- 0.404 vs 0.655 +/- 0.373, P = 0.0103). Colon cancer derived cell lines including DLD-1, Caco-2, and Lovo also expressed high levels of TTYH2. In contrast, transfection with siRNA-TTYH2 significantly inhibited both proliferation and scattering of these cancer cell lines.
The present work demonstrates, for the first time, that the TTYH2 gene expression is significantly up-regulated in colon cancer. The TTYH2 gene may play an important role in regulating both proliferating and metastatic potentials of colorectal cancer.
研究TTYH2在人结肠癌及结肠癌细胞系中的表达模式，并评估小干扰RNA（siRNA）对结肠癌细胞系中TTYH2表达的抑制作用。
我们通过逆转录聚合酶链反应（RT-PCR）研究TTYH2在结肠癌、癌旁非肿瘤性结肠黏膜及癌细胞系（DLD-1、caco-2和Lovo）中的表达模式。此外，构建针对TTYH2的siRNA质粒表达载体，并用Lipofectamine 2000转染至DLD-1和Caco-2细胞。通过RT-PCR检测TTYH2表达的下调情况，并通过MTT法和聚集试验评估siRNA在诱导细胞增殖和细胞聚集方面的作用。
与正常结肠黏膜相比，结肠癌组织中TTYH2基因表达显著上调（1.23±0.404对0.655±0.373，P = 0.0103）。包括DLD-1、Caco-2和Lovo在内的结肠癌衍生细胞系也高表达TTYH2。相反，用siRNA-TTYH2转染可显著抑制这些癌细胞系的增殖和分散。
本研究首次证明，TTYH2基因表达在结肠癌中显著上调。TTYH2基因可能在调节结直肠癌的增殖和转移潜能中起重要作用。
