The Role of STAT-3 in the Induction of Apoptosis in Pancreatic Cancer Cells by Benzyl Isothiocyanate
The Role of STAT-3 in the Induction of Apoptosis in Pancreatic Cancer Cells by Benzyl Isothiocyanate
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DOI：
10.1093/jnci/djn470
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发表时间：
2009-02-04
影响因子：
10.3
通讯作者：
Srivastava, Sanjay K.
中科院分区：
文献类型：
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作者：
Sahu, Ravi P.;Srivastava, Sanjay K.
异硫氰酸苄酯（BITC）是在十字花科蔬菜中发现的化合物，据报道具有抗癌特性，但其抑制人胰腺癌细胞生长的机制尚不完全清楚。人胰腺癌细胞（BxPC-3、AsPC-1、Capan-2、MiaPaCa-2和Panc-1）和永生化人胰腺细胞（HPDE-6）用载体或5-40 μ M的BITC处理，通过磺酰罗丹明B测定法评价细胞存活，通过半胱天冬酶-3和poly-ADP核糖聚合酶切割或通过商业细胞死亡测定法评价细胞凋亡。总的和激活的信号转导和转录激活因子-3（STAT-3）的蛋白表达的细胞进行了检查，通过蛋白质印迹，STAT-3的mRNA水平的逆转录-聚合酶链反应，和STAT-3的DNA结合和转录活性的市售的结合和报告分析。在无胸腺裸鼠中的BxPC-3胰腺肿瘤细胞异种移植物中研究了BITC治疗对肿瘤生长、凋亡和STAT-3蛋白表达的体内影响。BITC处理降低了BxPC-3、AsPC-1、Capan-2和MiaPaCa-2细胞的细胞存活率并诱导了细胞凋亡，并且在Panc-1细胞中的程度要小得多，但在HPDE-6细胞中没有。它还降低了激活的STAT-3蛋白和总STAT-3蛋白的水平，从而降低了STAT-3 DNA结合和转录活性。STAT-3在BxPC-3细胞中的过表达抑制BITC诱导的凋亡并恢复STAT-3活性。在喂食BITC的小鼠中，（60 μ mol/wk，5只小鼠，每组10个肿瘤），与对照小鼠相比，BxPC-3胰腺肿瘤异种移植物的生长受到抑制（6周时，对照小鼠与BITC处理小鼠的平均肿瘤体积= 334 vs 172 mm（3），差异=162 mm（3），95%置信区间= 118至204 mm（3）; P = 0.008）并且肿瘤具有增加的细胞凋亡和减少的STAT-3蛋白表达。BITC通过抑制STAT-3信号通路诱导某些类型的胰腺癌细胞的细胞凋亡。
Benzyl isothiocyanate (BITC), a compound found in cruciferous vegetables, has been reported to have anticancer properties, but the mechanism whereby it inhibits growth of human pancreatic cancer cells is incompletely understood.Human pancreatic cancer cells (BxPC-3, AsPC-1, Capan-2, MiaPaCa-2, and Panc-1) and immortalized human pancreatic cells (HPDE-6) were treated with vehicle or with BITC at 5-40 mu M, cell survival was evaluated by sulforhodamine B assay, and apoptosis by caspase-3 and poly-ADP ribose polymerase cleavage or by a commercial assay for cell death. Total and activated signal transducer and activator of transcription-3 (STAT-3) protein expression in the cells were examined by western blotting, STAT-3 mRNA levels by reverse transcription-polymerase chain reaction, and STAT-3 DNA-binding and transcriptional activity by commercially available binding and reporter assays. The effects of BITC treatment on tumor growth, apoptosis, and STAT-3 protein expression in vivo were studied in xenografts of BxPC-3 pancreatic tumor cells in athymic nude mice. All statistical tests were two-sided.BITC treatment reduced cell survival and induced apoptosis in BxPC-3, AsPC-1, Capan-2, and MiaPaCa-2 cells, and to a much lesser extent in Panc-1 cells, but not in HPDE-6 cells. It also reduced levels of activated and total STAT-3 protein, and as a result, STAT-3 DNA-binding and transcriptional activities. Overexpression of STAT-3 in BxPC-3 cells inhibited BITC-induced apoptosis and restored STAT-3 activity. In mice that were fed BITC (60 mu mol/wk, five mice, 10 tumors per group), growth of BxPC-3 pancreatic tumor xenografts was suppressed compared with control mice (at 6 weeks, mean tumor volume of control vs BITC-treated mice = 334 vs 172 mm(3), difference =162 mm(3), 95% confidence interval = 118 to 204 mm(3); P = .008) and tumors had increased apoptosis and reduced STAT-3 protein expression.BITC induces apoptosis in some types of pancreatic cancer cells by inhibiting the STAT-3 signaling pathway.