| 王连青,刘 剑,钱文斌.雷公藤内酯醇对肝癌细胞株HepG2的影响及作用机制[J].肿瘤学杂志,2013,19(12):959-963. |
| 雷公藤内酯醇对肝癌细胞株HepG2的影响及作用机制 |
| The Effect and Mechanism of Triptolide on Human Liver Cancer Cell Line HepG2 |
| 投稿时间：2013-11-12 |
| DOI：10.11735/j.issn.1671-170X.2013.12.B011 |
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| 中文关键词: 雷公藤内酯醇（TPL） 细胞凋亡 肝肿瘤 HepG2 |
| 英文关键词:triptolide（TPL） apoptosis liver neoplasms HepG2 |
| 基金项目: |
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| 中文摘要: |
| 摘 要：［目的］ 探讨雷公藤内酯醇（TPL）对肝癌细胞株HepG2的增殖抑制和凋亡诱导作用及作用机制。［方法］ MTT法检测TPL对细胞增殖的影响，Hoechst荧光染色观察凋亡细胞形态变化，AnnexinV/PI流式细胞仪标记法检测TPL对HepG2细胞凋亡作用的影响，Western blot检测caspase-3、8、9与PARP的表达情况。［结果］ 经MTT检测，10、20、40、80nmol/L浓度TPL作用24h后HepG2细胞的抑制率分别为15.2%、22.5%、34.9%和40.7%；48h后抑制率分别为21.5%、34.8%、44.1%和44.8%；72h后抑制率分别为31.6%、47.2%、51.6%和56.0%；抑制作用呈时间和剂量的依赖性。10、20、40、80nmol/L浓度的TPL作用24h的HepG2细胞，hoechst荧光染色显微镜下均出现明显的凋亡细胞；AnnexinV/PI流式细胞仪检测其凋亡率分别为8.89%、9.81%、14.69%和22.04%，对照组凋亡率为7.35%；Western blot结果显示，其caspase-3、9、PARP蛋白活化显影，且均较对照组显影增强，显影强度随TPL浓度的增加而增强，未见caspase-8活化显影。［结论］ TPL通过诱导HepG2凋亡发挥增殖抑制作用，其机制可能与caspase-3、9活化有关，即与线粒体通路相关，与caspase-8即死亡受体通路无关。 |
| 英文摘要: |
| Abstract: ［Purpose］ To investigate the proliferation and apoptosis effect of TPL on human liver cancer cell line HepG2 and its mechanism.［Methods］ TPL was added into HepG2 cells with the concentration of 10，20，40，80nmol/L as the TPL treated groups，and the culture medium alone was added as the control groups. The inhibitory rates in the HepG2 cells were assessed by MTT assay. In the HepG2 cells which had cultured for 24h，the morphological characters were observed by hoechst staining，and the apoptosis was determined by AnnexinV/ PI flow cytometry. The expression of activated caspase 3，8，9 and PARP proteins was examined by Western blot.［Results］ The inhibitory rates of HepG2 cultured for 24h with 10，20，40，80nmol/L concentration were15.2%，22.5%，34.9% and 40.7%；for 48h，21.5%，34.8%，44.1% and 44.8%；for 72h，31.6%，47.2%，51.6% and 56.0% respectively，in dose and time dependent manners. When the HepG2 cells treated by TPL cultured for 24h，typical apoptotic cells were observed via hoechst staining. The apoptosis rates of the HepG2 cells treated by TPL with 10，20，40，80nmol/L concentration were 8.89%，9.81%，14.69% and 22.04% respectively. The expression of actived caspase-3，9 and PARP proteins in the cells treated with TPL were higher than that of the control group，and the expression increased with the concentration of TPL.［Conclusion］ TPL could inhibit the proliferation and induce the apoptosis of HepG2. The apoptosis mechanism of TPL on HepG2 cells might relate to the expression of activated caspase-3，9 and PARP proteins and the mitochondrial pathway，might not relate with caspase-8 and the death receptor pathway. |
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