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Silencing IDH-2 gene by siRNA-IDH-2 inhibits human small cell lung carcinoma growth
Authors:LU Jian-hong  CHEN Guo-jun  DONG Chang-lin  GUO Shao-wen  JIN Yi-jun
Affiliation:1.Department of Clinical Laboratory, Zhejiang Provincial Hospital of Chinese Armed Police Force, Jiaxing 314000, China;2.Department of Pathology, Shanghai Shuguang Hospital of Traditional Chinese Medicine, Shanghai 200130, China.
Abstract:AIM:To investigate the effect of silencing isocitrate dehydrogenase 2 (IDH-2) gene by small interfering RNA (siRNA) on the biological characteristics of human small cell lung cancer cell line NCI-H446. METHODS:IDH-2 expression was knocked down in human small cell lung cancer cell line NCI-H446 by siRNA-IDH-2. The expression level of IDH-2 was determined by real-time PCR and Western blotting. The cell proliferation was measured by CCK-8 assay, the protein expression of MAPK p42 was detected by Western blotting, and the cell cycle was analyzed by flow cytometry. The migration was observed using Transwell cell migration system. BALB/c nude mice were subcutaneously injected on the back with NCI-H446 cells transfected with siRNA-IDH-2/negative control siRNA or non-transfected cells to study the tumor growth. RESULTS:siRNA-IDH-2 remarkably down-regulated the expression of IDH-2 and MAPK p42 in the NCI-H446 cells. siRNA-IDH-2 inhibited both the proliferation and migration abilities of NCI-H446 cells, and the cell cycle was arrested in S phase as compared with negative control group. Additionally, the volume of xenograft tumors in siRNA-IDH-2 group was significantly decreased as compared with control group. CONCLUSION:siRNA-IDH-2 down-regulates the expression of IDH-2 in NCI-H446 cells, reduces the cell migration efficiency and inhibits the tumor growth in vitro and in vivo.
Keywords:Isocitrate dehydrogenase 2  RNA interference  Lung neoplasms  
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