咔唑生物碱HY-1诱导K562细胞凋亡作用的研究  

Apoptosis-inducing effect of carbazole alkaloid (HY-1) in human erythroleukemia K562 cells

作　　者：蔡颖[1] 蔡兵[1] 崔承彬[2] 张冬云[1] 韩冰[1] 王元国[3] 王敏伟[3] 

机构地区：[1]天津生物医药研究所,300384 [2]军事医学科学院毒物药物研究所 [3]沈阳药科大学

出　　处：《中华肿瘤杂志》2005年第8期457-460,共4页Chinese Journal of Oncology

基　　金：国家重点基础研究发展规划项目(G1998051113);国家杰出青年科学基金资助项目(39825126)

摘　　要：目的研究咔唑生物碱HY1对人慢性骨髓性红白血病K562细胞的增殖抑制和凋亡诱导作用,并探讨其作用机制。方法通过SRB法检测HY1对K562细胞体外增殖的影响;通过PI荧光染色法、罗丹明123染色法、AnnexinⅤPI染色法等流式细胞术和琼脂糖凝胶电泳法,观察HY1处理K562细胞后凋亡相关指标的变化;通过Westernblot检测凋亡相关蛋白的变化。结果HY1对K562细胞体外增殖的半数抑制浓度(IC50)为(29.05±0.90)μmol/L。随着HY1药物浓度的增加和作用时间的延长,SubG1期细胞明显增加,线粒体膜电位迅速下降,发生凋亡的细胞比例也显著增加,而且经琼脂糖凝胶电泳检测到了典型的DNALadder。Westernblot结果显示,细胞色素c(Cytc)表达明显增加;caspase9酶原被剪切,且活化程度呈时间依赖性;caspase3酶原及其作用底物PARP也发生了剪切;caspase8表达无明显变化。结论HY1能够以剂量时间依赖方式,通过线粒体激活途径,诱导K562细胞发生凋亡,并由此发挥其抗肿瘤活性。Objective To investigate apoptosis-inducing effect and its mechanisms of HY-1, a carbazole alkaloid, on hmnan erythroleukemia K562 cells. Methods Cell proliferation was detected by sulforhodamine B （SRB） assay after treated with HY-1 at indicated doses. Cell cycle analysis was performed by flow cytometry, mitochondria membrane voltage change was assessed by rhodamine 123 staining, annexin V-PI apoptosis detecting kit and DNA agarose gel electrophoresis were used to identify apoptosis- inducing effect of HY-1. The alterations of apoptosis-relating proteins were detected by Western blot. Results The IC50 of HY- 1 in K562 cells was （29.05±0.90）μmoL/L by SRB assay. HY-1 had significant apoptotic inducing effect on K562 cells in a dose-and timedependent manner as verified by appearance of Sub-G1 peak on histogram of flow cytometry analysis, reduction of mitochondria membrane voltage, appearance of double positive cell group in Annexin V-PI apoptosis detecting test, and remarkable DNA ladder. The expression of cytosolic cytochrome c was apparently increased. Pro-caspase-9, pro-caspase-3 and PARP were all cleaved to active segments. There was no change in the expression of caspase-8. Conclusion HY-1 exerts its anticancer activity through triggering apoptosis of K562 cells by mitochondriaactivating pathways.

关 键 词：咔唑生物碱 人慢性骨髓性红白血病K562细胞 细胞增殖 细胞抑制 细胞凋亡 K562细胞 凋亡作用 琼脂糖凝胶电泳法 Western BLOT检测 

分 类 号：R733.7[医药卫生—肿瘤] R284.2[医药卫生—临床医学]