摘要
目的探讨蝎毒多肽对慢性粒细胞白血病细胞株K562中Shh和Gli1表达的影响及其机制。方法体外培养K562细胞,分为蝎毒多肽10、20、40mg/L组(A1、A2、A3组)、伊马替尼2g/L组(B组)以及生理盐水组(C组)。采用CCK-8法检测K562细胞增殖活性,RT-PCR法检测BCR/ABL、Shh、Gli1的mRNA表达,Western blot法检测P210^(BCR/ABL)、Shh、Gli1的蛋白表达。结果与C组相比,A2、A3、B组细胞增殖活性下降,Shh、Gli1的mRNA和蛋白表达以及BCR/ABL mRNA、P210^(BCR/ABL)蛋白的表达均降低(P<0.05)。A2、A3组细胞增殖活性以及Shh、Gli1的mRNA和蛋白表达水平与B组比较差异无统计学意义(P>0.05)。结论蝎毒多肽可以减少K562细胞增殖以及BCR/ABL mRNA和P210^(BCR/ABL)蛋白的表达,并通过抑制Shh和Gli1的表达阻断Hedgehog通路的激活,进而抑制慢性粒细胞白血病的进展。
Objective To investigate the effect and underlying mechanism of the peptide extracted from scorpion venom（PESV）on the expressions of Shh and Gli1 in chronic myelocytic leukemia cell line K562.Methods The K562 cells were cultured in vitro and divided into five groups of A1（incubated with PESV 10 mg/L）,A2（incubated with PESV 20 mg/L）,A3（incubated with PESV 40 mg/L）,B（incubated with imatinib 2 g/L）and C（incubated with normal saline 20μl）.The proliferation of K562 cells was determined by CCK-8 assay.The mRNA expressions of BCR/ABL,Shh and Gli1 were measured by RT-PCR.The protein expressions of P210BCR/ABL,Shh and Gli1 were detected by Western blot.Results Compared with group C,the proliferation of K562 cells was decreased,and the mRNA and protein expressions of Shh and Gli1,mRNA expression of BCR/ABL and protein expression of P210BCR/ABL were all significantly downregulated in groups of A2,A3 and B（P〈0.05）.There was no statistical difference in the cell proliferation and the mRNA and protein expressions of Shh and Gli1 among groups of A2,A3 and B（P〉0.05）.Conclusion The PESV can reduce the proliferation of K562 cells and the expressions of BCR/ABL mRNA and P210BCR/ABL protein,and block the activation of Hedgehog pathway by downregulating the expressions of Shh and Gli1,which then inhibits the progression of chronic myelocytic leukemia.
作者
张伟锋
杨文华
杨向东
王兴丽
ZHANG Weifeng;YANG Wenhua;YANG Xiangdong(Department of Hemotology,First Affiliated Hospital,Tianjin University of Traditional Chinese Medicine,Tianjin 300381,CHINA)
出处
《江苏医药》
CAS
2018年第7期729-732,共4页
Jiangsu Medical Journal
基金
高等学校博士学科点专项科研基金(20121210110004)
关键词
蝎毒多肽
HEDGEHOG通路
慢性粒细胞白血病
Peptide extracted from scorpion venom
Hedgehog pathway
Chronic myelocytic leukemia
