Triptolide induces cell death in pancreatic cancer cells by apoptotic and autophagic pathways.
Triptolide induces cell death in pancreatic cancer cells by apoptotic and autophagic pathways.
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DOI：
10.1053/j.gastro.2010.04.046
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发表时间：
2010-08
期刊：
影响因子：
29.4
通讯作者：
Saluja AK
中科院分区：
文献类型：
--
作者：
Mujumdar N;Mackenzie TN;Dudeja V;Chugh R;Antonoff MB;Borja-Cacho D;Sangwan V;Dawra R;Vickers SM;Saluja AK
胰腺癌是人类最致命的恶性肿瘤之一，对目前的化疗具有耐药性。我们先前已经证明雷公藤内酯醇在体外抑制胰腺癌细胞的生长，并在体内阻止肿瘤生长。这项研究调查了雷公藤内酯醇杀死胰腺癌细胞的机制，这是以前没有研究过的。用雷公藤内酯醇处理细胞，并使用比色测定法测量存活率和半胱天冬酶-3活性。流式细胞仪检测Annexin V、碘化丙啶和吖啶橙子染色。免疫荧光法用于监测细胞色素c和LC 3蛋白的定位。Caspase-3、Atg 5和Beclin 1水平通过将细胞暴露于它们各自的siRNA而下调。我们发现，雷公藤内酯醇诱导MiaPaCa-2，Capan-1和BxPC-3细胞的凋亡和S2-013，S2-VP 10和Hs 766 T细胞的自噬。雷公藤内酯醇诱导的自噬具有促死亡效应，需要自噬特异性基因atg 5或beclin 1，并且与Akt/mTOR/p70 S6 K通路的失活和ERK 1/2通路的上调相关。抑制S2-013和S2-VP 10细胞中的自噬通过凋亡途径导致细胞死亡，而抑制自噬和凋亡两者拯救细胞死亡。这项研究首次表明，雷公藤内酯醇通过两种不同的途径杀死胰腺癌细胞。它在MiaPaCa-2、Capan-1和BxPC-3中诱导半胱天冬酶依赖性凋亡死亡，在转移性细胞系S2-013、S2-VP 10和Hs 766 T中诱导半胱天冬酶非依赖性自噬死亡，从而使其成为针对广谱胰腺癌的有吸引力的化疗剂。
Pancreatic adenocarcinoma, among the most lethal human malignancies, is resistant to current chemotherapies. We have previously shown that triptolide inhibits the growth of pancreatic cancer cells in vitro and prevents tumor growth in vivo. This study investigates the mechanism by which triptolide kills pancreatic cancer cells, which has not been previously studied. Cells were treated with triptolide and viability and caspase-3 activity were measured using colorimetric assays. Annexin V, propidium iodide and acridine orange staining were measured by flow cytometry. Immunofluorescence was used to monitor the localization of cytochrome c and LC3 proteins. Caspase-3, Atg5 and Beclin1 levels were downregulated by exposing cells to their respective siRNA. We show that triptolide induces apoptosis in MiaPaCa-2, Capan-1 and BxPC-3 cells and autophagy in S2-013, S2-VP10 and Hs766T cells. Triptolide-induced autophagy has a pro-death effect, requires autophagy-specific genes, atg5 or beclin1, and is associated with the inactivation of the Akt/mTOR/p70S6K pathway and the upregulation of the ERK1/2 pathway. Inhibition of autophagy in S2-013 and S2-VP10 cells results in cell death via the apoptotic pathway whereas inhibition of both autophagy and apoptosis rescues cell death. This study shows, for the first time, that triptolide kills pancreatic cancer cells by two different pathways. It induces caspase-dependent apoptotic death in MiaPaCa-2, Capan-1 and BxPC-3 and caspase-independent autophagic death in metastatic cell lines, S2-013, S2-VP10 and Hs766T, thereby making it an attractive chemotherapeutic agent against a broad spectrum of pancreatic cancers.
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