Oridonin induces apoptosis via PI3K/Akt pathway in cervical carcinoma HeLa cell line
Oridonin induces apoptosis via PI3K/Akt pathway in cervical carcinoma HeLa cell line
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DOI：
10.1111/j.1745-7254.2007.00667.x
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发表时间：
2007-11-01
影响因子：
8.2
通讯作者：
Zeng, Hai-tao
中科院分区：
文献类型：
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作者：
Hu, Hong-zhen;Yang, Yue-bo;Zeng, Hai-tao
目的：研究冬凌草二萜类化合物冬凌草苷对人宫颈癌HeLa细胞株的诱导凋亡作用。方法：采用Hoechst 33342染色法进行形态学分析、核凝聚和染色质断裂的监测。采用3-（4,5 -二甲基噻唑-(2)-基）- 2,5 -二苯基溴化四氮唑（MTT）测定法评估细胞活力。采用流式细胞术和Western blotting检测HeLa细胞株细胞凋亡及凋亡相关活化情况。结果：冬凌草甲素对HeLa细胞株的增殖具有剂量依赖性和时间依赖性。Oridonin处理下调了蛋白激酶B （Akt）的激活，forkhead box class O （FOXO）转录因子和糖原合成酶激酶3 （GSK3）的表达。Oridonin还诱导细胞色素c的释放，并激活caspase-3和多磷酸腺苷核糖聚合酶裂解。此外，caspase抑制剂Z-D(OMe)-E(OMe)-V-D(OMe)-FMK （z-DEVD-fmk）可阻止caspase-3的激活，并消除鸢尾草素诱导的细胞死亡。最后，oridonin处理HeLa细胞系下调凋亡抑制蛋白的表达。结论：冬凌草素诱导细胞凋亡涉及多种分子途径。Oridonin可能抑制HeLa细胞系中磷脂酰肌醇3-激酶（Akt， FOXO和GSK3）的组成性激活靶点，抑制增殖并诱导caspase依赖性凋亡。
Aim: To investigate the apoptosis-inducing effect of oridonin, a diterpenoid isolated from Rabdosia rubescens, in the human cervical carcinoma HeLa cell line. Methods: A morphological analysis, nuclear condensation, and fragmentation of chromatin were monitored using Hoechst 33342 staining. Cell viability was assessed using the 3-(4, 5-dimethylthiazol-(2)-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay. Cell apoptosis and the apoptosis-related activation in the HeLa cell line were evaluated by flow cytometry and Western blotting. Results: Oridonin suppressed the proliferation of the HeLa cell line in a dose- and time-dependent fashion. Oridonin treatment downregulated the activation of protein kinase B (Akt), the expression of forkhead box class O (FOXO) transcription factor, and glycogen synthase kinase 3 (GSK3). Oridonin also induced the release of cytochrome c accompanied by the activation of caspase-3 and poly-adenosine diphosphate-ribose polymerase cleavage. In addition, Z-D(OMe)-E(OMe)-V-D(OMe)-FMK (z-DEVD-fmk), an inhibitor of caspases, prevented caspase-3 activation and abrogated oridonin-induced cell death. Finally, oridonin treatment of the HeLa cell line downregulated the expression of the inhibitor of the apoptosis protein. Conclusion: Our results showed that oridonin-induced apoptosis involved several molecular pathways. Oridonin may suppress constitutively activated targets of phosphatidylinositol 3-kinase (Akt, FOXO, and GSK3) in the HeLa cell line, inhibiting the proliferation and induction of caspase-dependent apoptosis.