蝎毒多肽对慢性粒细胞白血病细胞Hh通路下游活化因子Gli1表达的影响

摘要

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 .%目的 探讨蝎毒多肽对慢性粒细胞白血病细胞Hedgehog通路(Hh通路)下游活化因子Gli1表达的影响.方法 将慢性粒细胞白血病细胞K562接种于24孔板,随机分为蝎毒多肽1、2、3组和伊马替尼组、空白对照组.蝎毒多肽1、2、3组分别加入10、20、40μg/mL的蝎毒多肽20μL,伊马替尼组加入2 mg/mL的伊马替尼20μL,空白对照组加入生理盐水20μL,继续培养48 h.采用实时荧光定量PCR法检测各组Gli1 mRNA相对表达量,Western blotting法检测各组Gli1蛋白相对表达量.结果 蝎毒多肽1、2、3组和伊马替尼组、空白对照组Gli1 mRNA相对表达量分别为1.983±0.091、1.355±0.093、1.279±0.105、1.302±0.074、2.745±0.188;Gli1蛋白相对表达量分别为0.902±0.057、0.722±0.074、0.693±0.097、0.701±0.067、0.981±0.063.蝎毒多肽1组Gli1 mRNA和蛋白相对表达量与空白对照组比较P均>0.05;蝎毒多肽2、3组及伊马替尼组Gli1 mRNA和蛋白相对表达量明显低于蝎毒多肽1组及空白对照组(P均<0.05);蝎毒多肽2、3组及伊马替尼组Gli1 mRNA和蛋白相对表达量比较P均>0.05.结论 20～40μg/mL的蝎毒多肽可明显抑制慢性粒细胞白血病细胞Gli1基因表达.