Inhibitory effect of siRNA-Annexin A7 on growth, migration, and invasion in BGC823 cells and gastric cancer xenograftsin nude mice.

ABSTRACT

OBJECTIVE: To explore the inhibitory effect of siRNA-Annexin A7 on growth, migration, and invasion of transplanted gastric cancer in nude mice. METHODS: The siRNA sequence targeting to human Annexin A7 gene was designed, and based on that a pair of complementary oligonucleotides were synthesized, annealed, and cloned into plasmid pGenesil-1.1 to construct recombinant plasmid siRNA-Annexin A7. Transplanted gastric cancer model was established by injecting s.c. nude mice with human gastric cancer BGC823 cells, and siRNA-Annexin A7 was injected into the tumors formed. The nude mice were observed for clinical manifestation relying on the size and weight of transplanted tumors. The tumor tissue and angiogenesis were examined by pathologic sections. Flow cytometry was used to detect the changes of cell cycle. Western blot and qRT-PCR were used to analyze the expression of PCNA, P27, MMP-2, and TIMP-2. RESULTS: Both the size and weight of transplanted tumors of nude mice injected with siRNA-Annexin A7 were less than those of control groups (P<0.05). The examination of pathologic sections showed that, compared with in the control group, obvious necrosis of tumor cells was observed in siRNA-Annexin A7 group. The cells in stage S were fewer in siRNA-Annexin A7 group than those in the other two groups, while the cells in stage G0/G1 were much more in siRNA-Annexin A7 group. The results of western blot and qRT-PCR confirmed that the expression of PCNA and MMP-2 was down-regulated, whereas the expression of p27 was up-regulated. CONCLUSION: Gastric cancer xenografts were established in nude mice with human gastric cancer BGC823 cells. The volume and weight of tumor were decreased after inhibition of Annexin A7 expression in BGC823 cells. Tumor cells were arranged sparsely after inhibition of Annexin A7 expression in BGC823 cells. The siRNA-Annexin A7 inhibits Annexin A7 expression in transplanted gastric cancer of nude mice, and influences the growth, migration, and invasion of tumors by down-regulating the expression of PCNA and MMP-2, as well as up-regulating the expression of p27.