靶向抑制ERG的小分子多肽通过下调c-Myc杀伤急性淋巴性白血病细胞的机制研究

Study on the Mechanism of Killing Acute Lymphoblastic Leukemia Cells by Small Molecule Peptides Targeting ERG that Inhibit C-Myc Expression

关键词:  小分子多肽  ERG  c-Myc  急性淋巴细胞性白血病

Key words:  small molecule peptide  ERG  c-Myc  acute lymphoblastic leukemia

摘要:

目的 研究靶向抑制ERG的小分子多肽杀伤急性淋巴性白血病(acute lymphoblastic leukemia，ALL)细胞的机制。方法 CellTiter-Glo^®细胞活力测定试剂盒检测不同浓度的小分子多肽对ERG阳性的ALL细胞系增殖和活力的影响；Annexin V/PI双染法检测小分子多肽对ERG阳性的ALL细胞系的凋亡诱导情况；裸鼠皮下瘤模型用于评估小分子多肽对ERG阳性的ALL细胞系在体内杀伤作用；Western blotting检测蛋白酶体抑制剂对小分子多肽降解ERG蛋白的拮抗作用；慢病毒包装的质粒转染ALL细胞系，干扰ERG的表达，观察其对c-Myc的影响；慢病毒包装的质粒转染ALL细胞系沉默c-Myc的表达，观察其对筑巢因子GDF15表达的影响。结果 靶向降解ERG的小分子多肽在体内外模型中均可以杀伤ALL细胞系；在质粒转染敲低ERG及c-Myc表达的细胞系，GDF15表达被明显抑制。结论 本研究明确了小分子多肽在体外和体内具有杀伤ERG阳性ALL细胞的作用，其通过蛋白酶体降解的方式抑制ERG的表达，并可能通过c-Myc来抑制筑巢因子GDF15的表达，从而发挥杀伤ERG阳性的ALL细胞系的作用。

Abstract:

OBJECTIVE To study the mechanism of killing acute lymphoblastic leukemia(ALL) cells by targeting small molecule peptides that inhibit ERG.METHODS CellTiter-Glo^® cell viability assay kit was used to detect the effect of different concentrations of small molecule polypeptides on the proliferation and viability of ERG-positive ALL cell lines; Annexin V/PI double staining method was used to detect the apoptosis induction of molecular polypeptides on ERG-positive ALL cell lines. The nude mouse subcutaneous tumor model was used to evaluate the killing effect of small-molecule polypeptides on ERG-positive ALL cell lines in vivo; Western blotting detected the proteasome inhibitors on the degradation of ERG by small-molecule polypeptides; lentiviral-packaged plasmid transfected into ALL cell lines, which interfered with the expression of ERG, and observed its effect on c-Myc; lentiviral-packaged plasmid transfected into ALL cell lines to silence c-Myc expression, and observed its effect on nesting factor GDF15 expression. RESULTS Small molecule peptides targeting ERG degradation could kill ALL cell lines in vitro and in vivo; the expression of GDF15 was significantly inhibited in the cell lines that knocked down the expression of ERG and c-Myc by plasmid transfection. CONCLUSION This study clarifies that small molecule polypeptides have the effect of killing ERG-positive ALL cells in vitro and in vivo. They inhibit the expression of ERG through proteasomal degradation, and may inhibit the nesting factor GDF15 through c-Myc, thereby killing ERG-positive ALL cell lines.