Effects of PESV on the Hedgehog pathway downstream activating factor Gli1 expression of CML K562 cells

ZHANG Weifeng; YANG Wenhua

doi:10.3969/j.issn.1002-266X.2018.20.007

Abstract：Objective To investigate the effects of polypeptide extract from scorpion venom ( PESV) on the Hedgehog (Hh) pathway downstream activating factor Gli1 expression of CML K562 cells.Methods The cultured K562 cells were inoculated in 24-well plate, and then the cells were divided into the following five groups:PESV group 1, PESV group 2, PESV group 3, imatinib group and the blank control group .The cells in the PESV groups 1, 2, and 3 were added with 10, 20, and 40μg/mL PESV 20μL, respectively, cells in imatinib group were added with 2 mg/mL imatinib 20μL, and cells in the blank control group were added with saline 20 μL.At 48 h, the expression level of Gli1 mRNA was determined by PT-PCR, and the expression of Gli1 protein was determined by Western blotting .Results The relative expression levels of Gli1 mRNA in the PESV group 1, PESV group 2, PESV group 3, imatinib group, and the blank control group were 1.983 ±0.091, 1.355 ±0.093, 1.279 ±0.105, 1.302 ±0.074, and 2.745 ±0.188, respectively;the relative expression lev-els of Gli1 protein in each group were 0.902 ±0.057, 0.722 ±0.074, 0.693 ±0.097, 0.701 ±0.067, and 0.981 ± 0.063, respectively.There was no significant difference in the expression of Gli 1 mRNA and protein between the PESV group 1 and the blank control group (all P>0.05), the relative expression levels of Gli1 mRNA and protein in PESV group 2, PESV group 3 and imatinib group were significantly lower than those of the PESV group 1 and the blank control group (all P<0.05), but there was no significant difference among PESV group 2, PESV group 3, and imatinib group (all P>0.05).Conclusion PESV [(20-40) μg/mL] can obviously inhibit the expression of Gli1 gene in K562 cells, and Gli1 gene is a target for the treatment of CML by PESV .