Inhibition of tumor growth of nonª²small cell lung cancer by small interfering RNA targeting to HER2/neu gene

    REN Xinª²Ling£¬ QIAN Guiª²Sheng£¬ BAO Wei£¬ FU Haiª²Jing, JIA Linª²Tao£¬ ZHANG Rui£¬ WANG Tao£¬ XU Yanª²Ming£¬ YANG Anª²Gang

    Chinese PLA Institute of Respiratory Disease£¬ Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China, Department of Immunology, Department of Biochemistry and Molecular Biology, School of Basic Medicine, Fourth Military Medical University, Xi¬ðan 7l0033, China

    ¡¾Abstract¡¿ AIM: To construct RNA interference (RNAi) vector to downª²regulate HER2/neu gene and study the RNAi effect on the cell cycle and tumor growth of nonª²small cell lung cancer. METHODS: Oligonucleotides of 64 base pairs for hairpin RNA targeting HER2/neu were designed, chemically synthesized and annealed, and cloned into pSUPER vector.After identified by restriction digestion£¬the right vectors were transiently transfected into SPCª²Aª²1 cells, a human lung adenocarcinoma cell line. The HER2 mRNA was detected by RTª²PCR, and the protein detected by Western blot and indirect immunofluorescence staining. FCM analysis and MTT method were applied to measure cell cycle and cell growth respectively. RESULTS: SPCª²Aª²1 cells had an overexpression of HER2. The vector of RNAi which can interfere HER2/neu gene was successfully constructed, and compared with parent cells, the vector can increase the number of cells in G0/G1 phase by 9.9%, and inhibit the tumor cell growth (P<0.05). CONCLUSION: We successfully constructed an expressing hairpin RNA against HER2/neu vector. The vector can effectively silence HER2/neu gene, increase the number of cells in G0/G1 phase, and then slow down the growth of tumor cells.This may be a useful therapeutic strategy for NSCLC overª²expressing HER2/neu. ......