Down-regulation of telomerase activity and activation of caspase-3 are responsible for Tanshinone I-induced apoptosis in monocyte leukemia cells in vitro.
Down-regulation of telomerase activity and activation of caspase-3 are responsible for Tanshinone I-induced apoptosis in monocyte leukemia cells in vitro.
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端粒酶活性的下调和 caspase-3 的激活是丹参酮 I 诱导体外单核白血病细胞凋亡的原因。
DOI：
10.3390/ijms11062267
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发表时间：
2010-05-26
影响因子：
5.6
通讯作者：
Liu JJ
中科院分区：
文献类型：
--
作者：
Liu XD;Fan RF;Zhang Y;Yang HZ;Fang ZG;Guan WB;Lin DJ;Xiao RZ;Huang RW;Huang HQ;Liu PQ;Liu JJ
丹参酮I（Tanshinone I，Tan-I）是从传统草药丹参中提取的一种二萜醌类化合物。最近，Tan-I被报道具有抗肿瘤作用。本研究观察了Tan-I对三种单核细胞白血病细胞（U937、THP-1和SHI-1）的生长抑制和凋亡诱导作用。MTT法测定细胞活力。流式细胞术（FCM）和AnnexinV/PI染色检测细胞凋亡。采用逆转录聚合酶链反应（RT-PCR）和PCR-酶联免疫吸附法（ELISA）检测凋亡前后人端粒酶逆转录酶（hTERT）的表达和端粒酶活性。Caspase-3活性测定采用Caspase比色试剂盒和Western blot分析。采用ABI PRISM 7500序列检测系统，通过Western blot和Real-time RT-PCR检测凋亡抑制基因Survivin的表达。结果表明，Tan-I能抑制上述三种白血病细胞的生长，并诱导其凋亡，且呈时间和剂量依赖性。经Tan-I处理48 h后，Western blotting显示caspase-3酶原蛋白被切割，出现17 kD亚基，并清楚地发现caspase-3底物聚ADP核糖聚合酶（PARP）的89 kD切割产物。hTERTmRNA表达和端粒酶活性均呈剂量依赖性下降。Real-time RT-PCR和Western blot结果显示Survivin表达明显下调。提示Tan-I诱导单核细胞白血病U937、THP-1和SHI-1细胞凋亡与caspase-3的激活、hTERTmRNA表达和端粒酶活性的降低以及Survivin表达的下调密切相关。据我们所知，这是第一个关于Tan-I对单核细胞白血病细胞作用的报道。
Tanshinone I (Tan-I) is a diterpene quinone extracted from the traditional herbal medicine Salvia miltiorrhiza Bunge. Recently, Tan-I has been reported to have anti-tumor effects. In this study, we investigated the growth inhibition and apoptosis inducing effects of Tan-I on three kinds of monocytic leukemia cells (U937, THP-1 and SHI 1). Cell viability was measured by MTT assay. Cell apoptosis was assessed by flow cytometry (FCM) and AnnexinV/PI staining. Reverse transcriptase polymerase chain reaction (RT-PCR) and PCR–enzyme-linked immunosorbent assay (ELISA) were used to detect human telomerase reverse transcriptase (hTERT) expression and telomerase activity before and after apoptosis. The activity of caspase-3 was determined by Caspase colorimetric assay kit and Western blot analysis. Expression of the anti-apoptotic gene Survivin was assayed by Western blot and Real-time RT-PCR using the ABI PRISM 7500 Sequence Detection System. The results revealed that Tan-I could inhibit the growth of these three kinds of leukemia cells and cause apoptosis in a time- and dose-dependent manner. After treatment by Tan-I for 48 h, Western blotting showed cleavage of the caspase-3 zymogen protein with the appearance of its 17-kD subunit, and a 89-kD cleavage product of poly (ADP-ribose) polymerase (PARP), a known substrate of caspase-3, was also found clearly. The expression of hTERT mRNA as well as activity of telomerase were decreased concurrently in a dose-dependent manner. Moreover, Real-time RT-PCR and Western blot revealed a significant down-regulation of Survivin. We therefore conclude that the induction of apoptosis by Tan-I in monocytic leukemia U937 THP-1 and SHI 1 cells is highly correlated with activation of caspase-3 and decreasing of hTERT mRNA expression and telomerase activity as well as down-regulation of Survivin expression. To our knowledge, this is the first report about the effects of Tan-I on monocytic leukemia cells.
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