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Down-regulation of telomerase activity and activation of caspase-3 are responsible for tanshinone I-induced apoptosis in monocyte leukemia cells in vitro.
MedLine Citation:
PMID:  20640151     Owner:  NLM     Status:  In-Data-Review    
Abstract/OtherAbstract:
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.
Authors:
Xiao-Dan Liu; Rui-Fang Fan; Yong Zhang; Hong-Zhi Yang; Zhi-Gang Fang; Wei-Bing Guan; Dong-Jun Lin; Ruo-Zhi Xiao; Ren-Wei Huang; He-Qing Huang; Pei-Qing Liu; Jia-Jun Liu
Publication Detail:
Type:  Journal Article     Date:  2010-05-26
Journal Detail:
Title:  International journal of molecular sciences     Volume:  11     ISSN:  1422-0067     ISO Abbreviation:  Int J Mol Sci     Publication Date:  2010  
Date Detail:
Created Date:  2010-07-19     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101092791     Medline TA:  Int J Mol Sci     Country:  Switzerland    
Other Details:
Languages:  eng     Pagination:  2267-80     Citation Subset:  -    
Affiliation:
Hematological Department & Institute, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong 510630, China; E-Mails: lenovo381@126.com (X.-D.L.); xfangcn@163.com (R.-F.F.); fzg92@163.com (Z.-G.F.); dongjunlin0168@163.com (D.-J.L.); ruozhi_xiao@yahoo.com (R.-Z.X.); huangrw56@163.com (R.-W.H.).
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