[Lentiviral vector-mediated Nanog short-hairpin RNA inhibits the growth of human gastric carcinoma xenograft in nude mice].

OBJECTIVE

To construct lentiviral vector that interferes with Nanog, investigate its expression in human gastric cancer cell SGC-7901-transplanted nude mice, and explore the effect of shRNA-Nanog transfection on the growth of the xenograft in mice.

METHODS

Lentivirus carrying shRNA-Nanog was prepared by incorporating previously validated siRNA into Nanog gene specific lentiviral vectors. The models of human gastric cancer transplantation were constructed in nude mice. The mouse models were randomly divided into lentivirus-shRNA-Nanog transfection group (experimental group), GFP infection group (empty vector group) and uninfected group (control group). The tumor volume and mass changes were measured. Small animal in vivo imaging was employed to examine the fluorescent intensity and tumor metastasis. The Nanog protein expression was determined by Western blot analysis. The apoptosis of transinfected tumor cells was analyzed by TUNEL method. Results The gene sequencing demonstrated that the recombinant lentivirus carrying shRNA-Nanog was successfully established. In vivo imaging showed that 27 days after transfection, the total fluorescent intensity in the experimental group was significantly lower than that in the empty vector group or the uninfected group. The xenograft volume and mass in the experimental group decreased significantly as compared with those in the empty vector group or the uninfected group. Western blotting indicated that the expression of Nanog protein in the experimental group was significantly lower than that in the empty vector or uninfected group. TUNEL results revealed that the apoptosis rate in the experimental group was significantly higher than that in the other two groups. No statistically significant difference was found between the empty vector group and the uninfected group.

CONCLUSION

We successfully established Nanog gene-shRNA expression vector and capsulated it as lentivirus particles, which could inhibit the growth of xenograft in nude mice.