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Int. J. Mol. Sci. 2010, 11(6), 2267-2280; https://doi.org/10.3390/ijms11062267

Down-Regulation of Telomerase Activity and Activation of Caspase-3 Are Responsible for Tanshinone I-Induced Apoptosis in Monocyte Leukemia Cells in Vitro

1
Hematological Department & Institute, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong 510630, China
2
Department of Nuclear Medicine, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong 510630, China
3
Department of Traditional Chinese Medicine, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong 510630, China
4
Lab of Pharmacology and Toxicology, School of Pharmaceutical Science of Sun Yat-Sen University, Guangzhou, Guangdong 510080, China
These authors contributed equally to this work.
*
Author to whom correspondence should be addressed.
Received: 11 May 2010 / Accepted: 21 May 2010 / Published: 26 May 2010
(This article belongs to the Section Biochemistry, Molecular and Cellular Biology)
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Abstract

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 resultsrevealed 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. View Full-Text
Keywords: Tanshinone I (Tan-I); telomerase; survivin; leukemia Tanshinone I (Tan-I); telomerase; survivin; leukemia
This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).
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Liu, X.-D.; Fan, R.-F.; Zhang, Y.; Yang, H.-Z.; Fang, Z.-G.; Guan, W.-B.; Lin, D.-J.; Xiao, R.-Z.; Huang, R.-W.; Huang, H.-Q.; Liu, P.-Q.; Liu, J.-J. Down-Regulation of Telomerase Activity and Activation of Caspase-3 Are Responsible for Tanshinone I-Induced Apoptosis in Monocyte Leukemia Cells in Vitro. Int. J. Mol. Sci. 2010, 11, 2267-2280.

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