RNA干扰技术靶向P13K/AKT信号通路抵制胃腺癌SGC7901细胞侵袭转移的研究

Objective To observe the inhibitory effects on the invasion of gastric adenocarcinoma SGC7901 cells by small hairpin RNA targeting PIK3R1 and AKT1 in vitro. Methods The recombinant adenovirus vector plasmid expression vector which contained PIK3R1 and AKT1 shRNA was transfected into SGC7901 cells. Real time PCR and Western blot were used to detect the expression of P1K3R1 and AKT1. ELESA was used to measure the change in the MMP-2 and MMP-9 expression. The invasion ability of the tumor ceils was examined by Scarification, Transwell and 3-dimensional matrigel matrix tests. Re-sults rAdS-A-P mediated shRNA targeting PIK3R1 ,and AKT1 dramatically down-regulated their expres-sion in SGC7901 cells. MMP-2 and MMP-9 were downregulated,and TIMP-2 was upregulated. The extra-cellular levels of MMP-2 and MMP-9 were decreased. Scarification test indicated that the invision ability was decreased obviously. Transwell showed that the number of cells invading through the matrigel in con-trol. nonsense sequence, and rAd5-A-P transfection groups was 105.0±4.0,102.5±6.4, and 67.0±3.9 respectively. In the rAdS-A-P transfection group, cells formed only small aggregates as compared with the control and nonsense sequence groups in 3-dimensional matrigel matrix growth test. Conclusion ShRNA targeting PIK3R1 ,and AKTI downregulated significantly their expression in a sequence-specific manner, and inhibited the invasion of gastric adenocarcinoma SGC7901 cells in vitro.

Experimental study on targeting PDK/AKT signal pathway by RNAi suppressing gastric adenocarcinoma SGC7901 cell invasion in vitro