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Inhibitory effects of septin 2 gene silencing induced by siRNA on migration of human colorectal cancer cell line LoVo
Authors:DAI Xin-zhen  HUANG Xue-ping  ZHONG Lin  CHEN Shao-hong  HAN Xi-qun  ZHANG Yu  ZHU Mei-gang  ZHAO Tong
Institution:1. Department of Pathology, Nanfang Hospital, Guangzhou 510515, China;2. Department of Pathology, School of Basic Medical Science, Southern Medical University, Guangzhou 510515, China
Abstract:AIM:To investigate the expression of septin 2 in human colorectal cancer cell lines and the effect of septin 2 on the capacity of migration of human colorectal cancer cell line LoVo. METHODS:Real-time fluorescence quantitative RT-PCR and Western blotting analysis were used to determine the mRNA and protein levels of septin 2 in different metastatic potential cell lines, respectively. The expression of septin 2 in LoVo cells was silenced by siRNA. The efficacy of siRNA was confirmed by real-time fluorescence quantitative RT-PCR. Septin 2 and its co-localization with F-actin were measured by immunofluorescence method. The migration of transfected cells was analyzed by scratch test. RESULTS:The expression of septin 2 in LoVo cells was significantly higher than that in low metastatic potential cell lines such as SW480, HCT-116 and HT-29 at both mRNA and protein levels. The mRNA expression of septin 2was successfully silenced in LoVo cells by siRNA. Cell wound closure rate was also decreased in septin 2 siRNA group as compared with control group(both P<0.05). The co-expression of septin 2 and F-actin formed the typical filamentous-granular organization, and down-regulation of septin 2 resulted in cell skeleton disturbance with less or shorter pseudopodium and decreased stress fibers. CONCLUSION:Septin 2 is highly expressed in LoVo cells. Partial deletion of septin 2 represses the capability of tumor cell migration.
Keywords:Septin 2  LoVo cells  Cell skeleton  
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