摘要
目的探讨小分子化合物PKF118—310对人慢性粒细胞白血病（CML）K562细胞凋亡的影响及其机制。方法用不同浓度PKF118—310处理K562细胞，用四甲基偶氮唑盐（MTY）法检测其对细胞增殖的抑制；免疫荧光法观察细胞核β-catenin／TCF转录复合物的形成；流式细胞术检测细胞凋亡；Westernblot检测caspase-3、caspase-8、XIAP、bcl-2、β—catenin、TCF、C—myc及cyclinDI的表达。结果PKF118-310能够抑制K562细胞生长，其对K562细胞在24、48、72h的半数抑制浓度（IC50）值分别为5．388、3．290、1．566μmol／L。细胞核内形成β—catenin／TCF转录复合物。分别用1．6、3．2μmol／L的PKF118-10作用于K562细胞48h后，经AnnexinV．FITC／碘化丙啶（PI）双标检测细胞凋亡率分别为（48．0±0．9）％、（80．2±1．2）％，均高于对照组的（1．2±0．6）％（均P〈0．05）。不同浓度PKF118—310处理K562细胞72h后，caspase-3及caspase-8蛋白表达均高于对照组（均P〈0．05），而XIAP、bcl-2、β—catenin、TCF、c-Tnyc及cyclinD1蛋白表达均低于对照组（均P〈0．05）。结论PKF118—310可抑制K562细胞增殖，并诱导其凋亡。
Objective To investigate the effect of small molecule antagonists PKFI18-310 on apoptosis of human chronic myeloid leukemia cell line K562 and its mechanism. Methods After the treatment of PKF118-310 at different concentration, the proliferation inhibition in K562 cells was detected by MTr, the β-catenin/TCF transcription complex in the nucleus was observed by immunofluorescence, the apoptosis was detected by flow cytometry, and the expressions of caspase-3, caspase-8, MAP, bcl-2, β-catenin, TCF, c-myc and cyclin D1 were detected by Western blot. Results PKFll8-310 inhibited the proliferation of K562 cells. The median inhibitory concentration （IC50） of PKF118-310-treated K562 cells for 24 h, 48 h and 72 h were 5.388, 3.290, 1.566 μmol/L, respectively. The β-catenin/TCF transcription complex in the nucleus was observed. After the treatment with 1.6 and 3.2 μmol/L PKF118-310 for 48 h, the apoptosis rates of K562 cells were （48.0±0.9） % and （80.2±1.2） %, respectively, and were higher than that in the control group [（1.2±0.6） %] （both P 〈 0.05）. After the treatment of K562 cells with PKF118-310 at different concentration for 72 h, the expression levels of caspase-3 and caspase-8 were increased （both P 〈 0.05）, and the expression levels of XIAP, bcl-2, 13-catenin, TCF, c-myc and cyclin D1 were significantly decreased compared with control group （all P 〈 0.05）. Conclusion PKF118-310 can inhibit K562 cells＇ proliferation, and induce the apoptosis.
出处
《白血病．淋巴瘤》
CAS
2016年第6期344-348,共5页
Journal of Leukemia & Lymphoma
