Abstract:

OBJECTIVE:To investigate the effects on HCT116 cell proliferation and apoptosis by down-regulating PRPS2 gene expression. METHODS:PRPS2 gene interference vector sh-PRPS2 was constructed. The vector was not the transfected in normal control;the negative vector was transfected in negative control and the RNAi vector was transfected in experimental groups. The vectors were transfected to HCT116 cells after identified by DNA sequencing, and the level of PRPS2 mRNA and protein were detected by RT-PCR and Western blot, respectively. The ability of cell proliferation was evaluated by CCK8 kit. The cell cycle and apoptosis were assessed by flow cytometry. RESULTS:The RNAi expression vector of PRPS2 was successfully constructed as identified by DNA sequencing:sh-PRPS2-1, sh-PRPS2-2 and sh-PRPS2-3. The relative expression levels of PRPS2 mRNA were 0.61±0.03, 0.89±0.02 and 0.27±0.05 in HCT116 cells 72 h after transfected with sh-PRPS2-1, sh-PRPS2-2 and sh-PRPS2-3, respectively, all decreased significantly compared with control (P<0.05). The relative expression levels of PRPS2 protein were 0.37±0.06, 0.84±0.05 and 0.30±0.04, respectively, and also decreased significantly compared with control (P<0.05). 72 h after transfection, the inhibition rate of sh-PRPS2-3 proliferation was 19.8%±2.4%. The apoptosis rate of HCT116 cell was increased to 68.4%±4.6% 72 h post transfection. Sub-G1 population percentage was increased to 12.9%±3.8%. CONCLUSION: Down-regulation of PRPS2 expression level could effectively inhibit cell proliferation as well as promote apoptosis. HCT116 cell was arrested in G0/G1 phase. This study provided a research basis for tumor targeted therapy.

Key words: PRPS2, RNA interfere, HCT116 cell, colon cancer