Shiozaki Atsushi, Shimizu Hiroki, Ichikawa Daisuke, Konishi Hirotaka, Komatsu Shuhei, Kubota Takeshi, Fujiwara Hitoshi, Okamoto Kazuma, Iitaka Daisuke, Nakashima Shingo, Nako Yoshito, Liu Mingyao, Otsuji Eigo
Atsushi Shiozaki, Hiroki Shimizu, Daisuke Ichikawa, Hirotaka Konishi, Shuhei Komatsu, Takeshi Kubota, Hitoshi Fujiwara, Kazuma Okamoto, Daisuke Iitaka, Shingo Nakashima, Yoshito Nako, Eigo Otsuji, Division of Digestive Surgery, Department of Surgery, Kyoto Prefectural University of Medicine, Kyoto 602-8566, Japan.
World J Gastroenterol. 2014 Dec 21;20(47):17863-76. doi: 10.3748/wjg.v20.i47.17863.
To investigate the role of claudin 1 in the regulation of genes involved in cell migration and tumor necrosis factor alpha (TNF-α)-induced gene expression in human gastric adenocarcinoma cells.
Knockdown experiments were conducted with claudin 1 small interfering RNA (siRNA), and the effects on the cell cycle, apoptosis, migration and invasion were analyzed in human gastric adenocarcinoma MKN28 cells. The gene expression profiles of cells were analyzed by microarray and bioinformatics.
The knockdown of claudin 1 significantly inhibited cell proliferation, migration and invasion, and increased apoptosis. Microarray analysis identified 245 genes whose expression levels were altered by the knockdown of claudin 1. Pathway analysis showed that the top-ranked molecular and cellular function was the cellular movement related pathway, which involved MMP7, TNF-SF10, TGFBR1, and CCL2. Furthermore, TNF- and nuclear frctor-κB were the top-ranked upstream regulators related to claudin 1. TNF-α treatment increased claudin 1 expression and cell migration in MKN28 cells. Microarray analysis indicated that the depletion of claudin 1 inhibited 80% of the TNF-α-induced mRNA expression changes. Further, TNF-α did not enhance cell migration in the claudin 1 siRNA transfected cells.
These results suggest that claudin 1 is an important messenger that regulates TNF-α-induced gene expression and migration in gastric cancer cells. A deeper understanding of these cellular processes may be helpful in establishing new therapeutic strategies for gastric cancer.
研究紧密连接蛋白1（claudin 1）在调控人胃腺癌细胞中参与细胞迁移的基因及肿瘤坏死因子α（TNF-α）诱导的基因表达中的作用。
采用claudin 1小干扰RNA（siRNA）进行敲低实验，并分析其对人胃腺癌MKN28细胞的细胞周期、凋亡、迁移和侵袭的影响。通过基因芯片和生物信息学分析细胞的基因表达谱。
claudin 1的敲低显著抑制细胞增殖、迁移和侵袭，并增加凋亡。基因芯片分析鉴定出245个基因，其表达水平因claudin 1的敲低而改变。通路分析表明，排名靠前的分子和细胞功能是细胞运动相关通路，其中涉及基质金属蛋白酶7（MMP7）、肿瘤坏死因子配体超家族成员10（TNF-SF10）、转化生长因子β受体1（TGFBR1）和趋化因子配体2（CCL2）。此外，TNF和核因子κB是与claudin 1相关的排名靠前的上游调节因子。TNF-α处理增加了MKN28细胞中claudin 1的表达和细胞迁移。基因芯片分析表明，claudin 1的缺失抑制了80%的TNF-α诱导的mRNA表达变化。此外，TNF-α在claudin 1 siRNA转染的细胞中未增强细胞迁移。
这些结果表明，claudin 1是调节胃癌细胞中TNF-α诱导的基因表达和迁移的重要信使。对这些细胞过程的深入了解可能有助于建立新的胃癌治疗策略。
