Wang Pan, Zeng Yun, Liu Tao, Zhang Chao, Yu Pei-Wu, Hao Ying-Xue, Luo Hua-Xin, Liu Gang
Pan Wang, Department of General Surgery, Affiliated Hospital of North Sichuan Medical College, Nanchong 637007, Sichuan Province, China.
World J Gastroenterol. 2014 Feb 28;20(8):2071-8. doi: 10.3748/wjg.v20.i8.2071.
To investigate the mechanisms of chloride intracellular channel 1 (CLIC1) in the metastasis of colon cancer under hypoxia-reoxygenation (H-R) conditions.
Fluorescent probes were used to detect reactive oxygen species (ROS) in LOVO cells. Wound healing assay and transwell assay were performed to examine the migration and invasion of LOVO cells. Expression of CLIC1 mRNA and protein, p-ERK, MMP-2 and MMP-9 proteins was analyzed by reverse transcription-polymerase chain reaction and Western blot.
H-R treatment increased the intracellular ROS level in LOVO cells. The mRNA and protein expression of CLIC1 was elevated under H-R conditions. Functional inhibition of CLIC1 markedly decreased the H-R-enhanced ROS generation, cell migration, invasion and phosphorylation of ERK in treated LOVO cells. Additionally, the expression of MMP-2 and MMP-9 could be regulated by CLIC1-mediated ROS/ERK pathway.
Our results suggest that CLIC1 protein is involved in the metastasis of colon cancer LOVO cells via regulating the ROS/ERK pathway in the H-R process.
探讨氯离子细胞内通道1（CLIC1）在缺氧复氧（H-R）条件下结肠癌转移中的作用机制。
使用荧光探针检测LOVO细胞中的活性氧（ROS）。采用划痕愈合实验和Transwell实验检测LOVO细胞的迁移和侵袭能力。通过逆转录聚合酶链反应和蛋白质免疫印迹法分析CLIC1 mRNA和蛋白、p-ERK、MMP-2和MMP-9蛋白的表达。
H-R处理增加了LOVO细胞内的ROS水平。在H-R条件下，CLIC1的mRNA和蛋白表达升高。CLIC1的功能抑制显著降低了H-R增强的LOVO细胞中ROS的产生、细胞迁移、侵袭以及ERK的磷酸化。此外，MMP-2和MMP-9的表达可由CLIC1介导的ROS/ERK途径调节。
我们的结果表明，CLIC1蛋白通过在H-R过程中调节ROS/ERK途径参与结肠癌LOVO细胞的转移。
