Basic & Clinical Medicine ›› 2011, Vol. 31 ›› Issue (6): 609-614.

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Construction of recombinant lentiviral vector of MicroRNA-29b and its inhibition on the migration and proliferation of gastric cancer cells

Chong-liang JIANG1,Xiao-yan WANG1,Jia-nan GONG2,Fang WANG2,Jia YU3,Tao FENG1   

  1. 1. Molecular Medicine&Cancer Research Center, Institute of Basic Medical Sciences, Chongqing Medical University
    3. Institute of Basic Medical Sciences, CAMS&PUMC
  • Received:2011-02-28 Revised:2011-04-15 Online:2011-06-05 Published:2011-06-06
  • Contact: Tao FENG

Abstract: Objective: To construct the recombinant lentivirus vector expressing hsa-miR-29b and observe the effect of over-expression of miR-29b on gastric cancer cell migration and proliferation. Methods The pMIR-miR-29b lentivirus vector was generated by PCR amplification of miR-29b precursor sequence. 293TN cells were transfected with pMIR-miR-29b、Packaging Plasmid(s) and pVSV-G plasmids to package the recombinant lentivirus. HGC-27 cells were infected with recombinant lentivirus and the expression level of miR-29b was measured by real-time PCR.The expression of target gene MCL-1 was tested by western blot .Scratch test and CCK8 were used to observe the change in cell migration and proliferation after miR-29b was over-expressed in gastric cancer cells. Results The expression vector of 29b restructuring slow virus have been successfully achieved. After infected stomach cancer cell HGC-27, successfully achieved the overexpression of miR-27. The miR-29b that is overexpressed can inhibit the expression of target gene MCL-1 in HGC-27, and inhibit the ability of migration and proliferation of HGC-27. comprised with untreated group and Lenti-vector group, the migration ability of Lenti-29b group is reduced significantly (P<0.01); comprised with Lenti-vector group, the Proliferative capacity of lenti-29b group is reduced significantly at 48h,72h and 96h(P<0.01). Conclusion Restructuring lentivirus particles that has over expression of miR-29b have been successful achieved. MiR-29b can inhibit the expression of target gene MCL-1 and the migration and proliferation of HGC-27.

Key words: miR-29b, Gastric cancer cells, Lentivirus, Cell migration

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