蝎毒抗癌多肽对HL-60细胞凋亡及Bcl-2和p53基因表达的影响

目的：研究蝎毒抗癌多肽（APBMV）在体外诱导人急性粒细胞白血病HL-60细胞凋亡的作用及其相关机制。方法采用CCK-8法检测APBMV作用后HL-60细胞的增殖抑制率，光镜观察APBMV作用后HL-60细胞的形态学变化，流式细胞仪检测细胞凋亡率，Western blot检测APBMV对相关凋亡基因蛋白表达的影响。结果 CCK-8法显示APBMV能够抑制HL-60细胞增殖，其对HL-60细胞增殖的IC50为15.64μg/ml。流式细胞仪检测提示APBMV能诱导HL-60细胞凋亡，且呈浓度-效应关系。APBMV（16μg/ml）作用48h后，细胞凋亡率为（36.1±2.1）%，与正常对照组比较有统计学差异（P＜0.05）。Western blot提示APBMV （4、8、12和16μg/ml）组的Bcl-2蛋白表达量分别为正常对照组的78.60%、54.60%、25.50%和4.20%；p53蛋白表达量分别为正常对照组的163.00%、271.00%、355.00%和447.00%。结论 APBMV可有效抑制HL-60细胞的增殖，并诱导细胞凋亡，其机制可能与促进HL-60细胞内p53基因的表达，抑制Bcl-2基因的表达有关。

Effect of anticancer polypeptide in scorpion venom on apoptosis and Bcl-2, p53 gene expression in human leukemia HL-60 cells

Objective To investigate the effect of antitumor polypeptide extracted from scorpion venom （APBMV） on apoptosis and Bcl- 2, p53 gene expression in human leukemia HL- 60 cells. Methods APBMV was separated and purified by column chromatography with SephadexG- 50, and analyzed by HPLC method. The cultured HL- 60 cells were treated with differ-ent concentrations of APBMV. The cellgrowth was assayed by CCK- 8 method;the morphological changes of HL- 60 cells was observe by microscope, cellapoptosis was measured by flow cytometry, the expression of Bcl- 2 and p53 genes was detected by Western blot. Results CCK- 8 demonstrated that APBMV restrained HL- 60 cells multiplication in a dose- effect manner with a IC50 of 15.64μg/ml. Flow cytometer showed that APBMV induced HL- 60 cells apoptosis in a dose- effect manner;with APBMV （16μg/ml） for 48h, the apoptosis rate of HL- 60 cells was （36.1＆#177;2.1）%, significantly higher than that of control group. Western blot showed that APBMV down- regulated the expression of Bcl- 2 protein and up- regulated expression of p53 protein. Conclu-sion APBMV can effectively inhibit the proliferation and induce apoptosis of HL- 60 cells, which are associated with up- regula-tion of p53 and down- regulation of Bcl- 2 genes.