Department of Biochemistry, Jin Lin Medical College, Ji Lin 132013, P.R. China.
Can J Physiol Pharmacol. 2012 Nov;90(11):1553-8. doi: 10.1139/y2012-134. Epub 2012 Nov 18.
Induction of apoptosis in tumor cells has become the major focus of anti-tumor therapeutics development. Juglone, a major chemical constituent of Juglans mandshurica Maxim, possesses several bioactivities, including anti-tumor. In the present study, HeLa cells were incubated with juglone at various concentrations. The proliferation inhibition of juglone on HeLa cells was tested by the MTT assay. Occurrence of apoptosis was detected by Hoechst 33258 staining, flow cytometry, and transmission electron microscopy. The expression of apoptotic-related proteins was examined by Western blot. The results showed that juglone inhibits the growth of HeLa cells in dose-dependent manner. Topical morphological changes of apoptotic body formation after juglone treatment were observed. The percentages of early apoptosis of Annexin V-FITC were 5.23%, 7.95%, 10.69%, and 20.92% with the concentrations of juglone (12.5, 25, 50, and 100 µmol/L), respectively. After cells were treated with juglone at the different dose for 24 h, the expression of Bcl-2 was significantly down-regulated and the expression of Bax was significantly up-regulated compared with the control. These events paralleled with activation of caspase-9, -8, -3, and PARP cleavage. The results suggest that juglone may be effective for the treatment of HeLa cells.
诱导肿瘤细胞凋亡已成为抗肿瘤治疗发展的主要焦点。胡桃醌是胡桃科胡桃属植物的主要化学成分之一,具有多种生物活性,包括抗肿瘤活性。在本研究中,用不同浓度的胡桃醌孵育 HeLa 细胞。用 MTT 法检测胡桃醌对 HeLa 细胞的增殖抑制作用。用 Hoechst 33258 染色、流式细胞术和透射电镜检测细胞凋亡的发生。用 Western blot 检测凋亡相关蛋白的表达。结果表明,胡桃醌呈剂量依赖性抑制 HeLa 细胞的生长。经胡桃醌处理后观察到凋亡小体形成的典型形态学变化。用浓度为 12.5、25、50 和 100 μmol/L 的胡桃醌处理细胞后,早期凋亡的 Annexin V-FITC 分别为 5.23%、7.95%、10.69%和 20.92%。用不同剂量的胡桃醌处理细胞 24 h 后,与对照组相比,Bcl-2 的表达明显下调,Bax 的表达明显上调。这些事件与 caspase-9、-8、-3 的激活和 PARP 切割平行。结果表明,胡桃醌可能对 HeLa 细胞有效。