鬼臼毒素衍生物QW-83对人宫颈癌HeLa细胞凋亡的影响及其机制研究
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篇名: 鬼臼毒素衍生物QW-83对人宫颈癌HeLa细胞凋亡的影响及其机制研究
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摘要: 目的:研究鬼臼毒素衍生物QW-83对人宫颈癌HeLa细胞凋亡的影响及其机制。方法:以0(阴性对照)、0.01、0.1、1、10    μmol/L QW-83和阳性药物依托泊苷(VP-16)分别培养人宫颈癌HeLa细胞48 h后,采用MTT法测定细胞的增殖抑制率和半数抑制浓度(IC50);以0(阴性对照)、2.5、5、10 μmol/L QW-83培养细胞48 h后,采用Hochest 33342染色观察细胞形态,采用流式细胞术测定细胞凋亡率,采用半定量逆转录-聚合酶链反应(RT-PCR)法检测细胞中凋亡相关基因P53、Bax、Casepase-3、Casepase-8、Casepase-9和Bcl-2 mRNA的表达。结果:与阴性对照比较,1、10 μmol/L VP-16和QW-83对HeLa细胞增殖有明显的抑制作用(P<0.05或P<0.01),IC50值分别为(5.11±0.43)、(4.96±0.54) μmol/L;染色结果显示,QW-83作用后细胞凋亡明显,细胞固缩;流式细胞检测结果显示,QW-83作用后细胞凋亡率呈浓度依赖性升高,为16.89%~62.56%;RT-PCR结果显示,QW-83作用后细胞中P53、Bax、Casepase-3、Casepase-8、Casepase-9 mRNA的表达增强,Bcl-2 mRNA的表达减弱,Bax/Bcl-2比例升高(P<0.05)。结论:鬼臼毒素衍生物QW-83能诱导人宫颈癌HeLa细胞的凋亡,其机制可能与调控凋亡相关基因mRNA的表达有关。
ABSTRACT: OBJECTIVE: To study the effects of podophyllotoxin derivative QW-83 on human cervical cancer HeLa cell apoptosis and its mechanism. METHODS: After treated with 0(negative control), 0.01, 0.1, 1 and 10 μmol/L QW-83 and positive drug etoposide (VP-16) for 48 h, proliferation inhibition rate and IC50 of HeLa cell were determined by MTT assay. The morphological changes of HeLa cell were observed by Hochest 33342 staining after treated with QW-83 [0 (negative control), 2.5, 5, 10 μmol/L] for 48 h; flow cytometry was used to detect apoptosis rate; semi quantitative RT-PCR was adopted to detect the expression of apoptosis related gene P53, Bax, Casepase-3, Casepase-8, Casepase-9 and Bcl-2 mRNA. RESULTS: Compared with negative control, 1, 10 μmol/L VP-16 and QW-83 had obvious proliferation inhibition effect on HeLa cells (P<0.05 or P<0.01), and IC50 were (5.11±0.43) μmol/L and (4.96±0.54) μmol/L. Hochest 33342 staining results showed QW-83 could obviously induce cells apoptosis and nuclear pyknosis. Flow cytometry showed QW-83 could increase apoptosis rate in concentration-dependent manner, being 16.89%-62.56%. RT-PCR showed mRNA expression of P53, Bax, Caspase-3, Casepase-8 and Casepase-9, Bcl-2/Bax increased, while mRNA expression of Bcl-2 decreased after treated with QW-83 (P<0.05). CONCLUSIONS: Podophyllotoxin derivative QW- 83 can induce HeLa cell apoptosis, and its mechanism may be associated with regulate mRNA expression of apoptosis related gene.
期刊: 2016年27卷第7期
作者: 冷玲,高晨光,陈虹,牛聪,曹波
AUTHORS: LENG Ling,GAO Chenguang,CHEN Hong,NIU Cong,CAO Bo
关键字: 鬼臼毒素衍生物;QW-83;人宫颈癌HeLa细胞;抗肿瘤活性;凋亡相关基因;依托泊苷
KEYWORDS: Podophyllotoxin derivative; QW-83; Human cervical cancer HeLa cell; Antitumor activity; Apoptosis-related gene; Etoposide
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