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Isocryptotanshinone Induced Apoptosis and Activated MAPK Signaling in Human Breast Cancer MCF-7 Cells.

Zhang X, Luo W, Zhao W, Lu J, Chen X - J Breast Cancer (2015)

Bottom Line: Apoptosis was determined by Hoechst 33342 staining, DNA fragmentation assays, and Western blotting for apoptotic proteins.ICTS significantly inhibited proliferation of MCF-7 and MDA-MB-231 human breast cancer cells, HepG2 human liver cancer cells, and A549 human lung cancer cells in vitro.Hoechst 33342 staining and Western blot analysis for apoptotic proteins suggested that ICTS induced apoptosis in MCF-7 cells.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Macao, China.

ABSTRACT

Purpose: Isocryptotanshinone (ICTS) is a natural bioactive product that is isolated from the roots of the widely used medical herb Salvia miltiorrhiza. However, few reports exist on the mechanisms underlying the therapeutic effects of ICTS. Here, we report that ICTS has anticancer activity and describe the mechanism underlying this effect.

Methods: The antiproliferative effect of ICTS was determined using 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) and clonogenic assays. The effect of ICTS on the cell cycle was measured using flow cytometry. Apoptosis was determined by Hoechst 33342 staining, DNA fragmentation assays, and Western blotting for apoptotic proteins. Finally, the effect of ICTS on mitogen-activated protein kinases (MAPKs) was determined by Western blotting.

Results: ICTS significantly inhibited proliferation of MCF-7 and MDA-MB-231 human breast cancer cells, HepG2 human liver cancer cells, and A549 human lung cancer cells in vitro. Among the tested cell lines, MCF-7 cells showed the highest sensitivity to ICTS. ICTS significantly inhibited colony formation by MCF-7 cells. Furthermore, exposure of MCF-7 cells to ICTS induced cell cycle arrest at the G1 phase and decreased mitochondrial membrane potential. Hoechst 33342 staining and Western blot analysis for apoptotic proteins suggested that ICTS induced apoptosis in MCF-7 cells. In addition, ICTS activated MAPK signaling in MCF-7 cells by inducing time- and concentration-dependent phosphorylation of JNK, ERK, and p38 MAPK.

Conclusion: Our results suggest that ICTS inhibited MCF-7 cell proliferation by inducing apoptosis and activating MAPK signaling pathways.

No MeSH data available.


Related in: MedlinePlus

The chemical structure and cytotoxicity of isocryptotanshinone (ICTS). (A) The chemical structure of ICTS. (B) MCF-7, MDA-MB-231, HepG2, and A549 cells were treated with indicated concentrations (0-40 µM) of ICTS for 24 hours. Cell viabilities were measured by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay. (C) After treatment with ICTS (0-20 µM) for 24 hours, morphological changes of MCF-7 cells were photographed. (D) MCF-7 cells were treated with indicated concentrations of ICTS for 24 hours. At a concentration of 20 µM, ICTS almost completely inhibited colony formation by MCF-7 cells.
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Figure 1: The chemical structure and cytotoxicity of isocryptotanshinone (ICTS). (A) The chemical structure of ICTS. (B) MCF-7, MDA-MB-231, HepG2, and A549 cells were treated with indicated concentrations (0-40 µM) of ICTS for 24 hours. Cell viabilities were measured by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay. (C) After treatment with ICTS (0-20 µM) for 24 hours, morphological changes of MCF-7 cells were photographed. (D) MCF-7 cells were treated with indicated concentrations of ICTS for 24 hours. At a concentration of 20 µM, ICTS almost completely inhibited colony formation by MCF-7 cells.

Mentions: Isocryptotanshinone (ICTS) (Figure 1A) was first isolated from S. miltiorrhiza in 1969 [8]. However, there are few reports on the biological activities of ICTS, despite the established use of S. miltiorrhiza in traditional Chinese medicine. Han et al. [9] reported that ICTS and two other tanshinones noncompetitively inhibited the activity of protein tyrosine phosphatase 1B, indicating their potential as treatments for type 2 diabetes. In recent decades, tanshinones have been widely investigated for their anticancer effects [710]. Cryptotanshinone (CTS), an important bioactive constituent of S. miltiorrhiza, inhibits cancer cell proliferation and induces apoptosis in several types of cancer cells, including MCF-7 human breast cancer cells and HepG2 liver carcinoma cells [111213]. The structure of ICTS is similar to that of CTS. Herein, we report the results of our investigation of the anticancer activity of ICTS.


Isocryptotanshinone Induced Apoptosis and Activated MAPK Signaling in Human Breast Cancer MCF-7 Cells.

Zhang X, Luo W, Zhao W, Lu J, Chen X - J Breast Cancer (2015)

The chemical structure and cytotoxicity of isocryptotanshinone (ICTS). (A) The chemical structure of ICTS. (B) MCF-7, MDA-MB-231, HepG2, and A549 cells were treated with indicated concentrations (0-40 µM) of ICTS for 24 hours. Cell viabilities were measured by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay. (C) After treatment with ICTS (0-20 µM) for 24 hours, morphological changes of MCF-7 cells were photographed. (D) MCF-7 cells were treated with indicated concentrations of ICTS for 24 hours. At a concentration of 20 µM, ICTS almost completely inhibited colony formation by MCF-7 cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4490259&req=5

Figure 1: The chemical structure and cytotoxicity of isocryptotanshinone (ICTS). (A) The chemical structure of ICTS. (B) MCF-7, MDA-MB-231, HepG2, and A549 cells were treated with indicated concentrations (0-40 µM) of ICTS for 24 hours. Cell viabilities were measured by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay. (C) After treatment with ICTS (0-20 µM) for 24 hours, morphological changes of MCF-7 cells were photographed. (D) MCF-7 cells were treated with indicated concentrations of ICTS for 24 hours. At a concentration of 20 µM, ICTS almost completely inhibited colony formation by MCF-7 cells.
Mentions: Isocryptotanshinone (ICTS) (Figure 1A) was first isolated from S. miltiorrhiza in 1969 [8]. However, there are few reports on the biological activities of ICTS, despite the established use of S. miltiorrhiza in traditional Chinese medicine. Han et al. [9] reported that ICTS and two other tanshinones noncompetitively inhibited the activity of protein tyrosine phosphatase 1B, indicating their potential as treatments for type 2 diabetes. In recent decades, tanshinones have been widely investigated for their anticancer effects [710]. Cryptotanshinone (CTS), an important bioactive constituent of S. miltiorrhiza, inhibits cancer cell proliferation and induces apoptosis in several types of cancer cells, including MCF-7 human breast cancer cells and HepG2 liver carcinoma cells [111213]. The structure of ICTS is similar to that of CTS. Herein, we report the results of our investigation of the anticancer activity of ICTS.

Bottom Line: Apoptosis was determined by Hoechst 33342 staining, DNA fragmentation assays, and Western blotting for apoptotic proteins.ICTS significantly inhibited proliferation of MCF-7 and MDA-MB-231 human breast cancer cells, HepG2 human liver cancer cells, and A549 human lung cancer cells in vitro.Hoechst 33342 staining and Western blot analysis for apoptotic proteins suggested that ICTS induced apoptosis in MCF-7 cells.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Macao, China.

ABSTRACT

Purpose: Isocryptotanshinone (ICTS) is a natural bioactive product that is isolated from the roots of the widely used medical herb Salvia miltiorrhiza. However, few reports exist on the mechanisms underlying the therapeutic effects of ICTS. Here, we report that ICTS has anticancer activity and describe the mechanism underlying this effect.

Methods: The antiproliferative effect of ICTS was determined using 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) and clonogenic assays. The effect of ICTS on the cell cycle was measured using flow cytometry. Apoptosis was determined by Hoechst 33342 staining, DNA fragmentation assays, and Western blotting for apoptotic proteins. Finally, the effect of ICTS on mitogen-activated protein kinases (MAPKs) was determined by Western blotting.

Results: ICTS significantly inhibited proliferation of MCF-7 and MDA-MB-231 human breast cancer cells, HepG2 human liver cancer cells, and A549 human lung cancer cells in vitro. Among the tested cell lines, MCF-7 cells showed the highest sensitivity to ICTS. ICTS significantly inhibited colony formation by MCF-7 cells. Furthermore, exposure of MCF-7 cells to ICTS induced cell cycle arrest at the G1 phase and decreased mitochondrial membrane potential. Hoechst 33342 staining and Western blot analysis for apoptotic proteins suggested that ICTS induced apoptosis in MCF-7 cells. In addition, ICTS activated MAPK signaling in MCF-7 cells by inducing time- and concentration-dependent phosphorylation of JNK, ERK, and p38 MAPK.

Conclusion: Our results suggest that ICTS inhibited MCF-7 cell proliferation by inducing apoptosis and activating MAPK signaling pathways.

No MeSH data available.


Related in: MedlinePlus