Limits...
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

Effect of isocryptotanshinone (ICTS) on mitochondrial membrane potential (MMP). MCF-7 cells were treated with 20 µM ICTS for 0, 2, 4, 8, and 20 hours, the MMP was determined by flow cytometer using JC-1 staining. Cells treated with 10 µM carbonyl cyanide 3-chlorophenylhydrazone (CCCP) for 20 minutes were used as the positive control.*p<0.05; †p<0.01 compared with 0 hour treatment group.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4490259&req=5

Figure 4: Effect of isocryptotanshinone (ICTS) on mitochondrial membrane potential (MMP). MCF-7 cells were treated with 20 µM ICTS for 0, 2, 4, 8, and 20 hours, the MMP was determined by flow cytometer using JC-1 staining. Cells treated with 10 µM carbonyl cyanide 3-chlorophenylhydrazone (CCCP) for 20 minutes were used as the positive control.*p<0.05; †p<0.01 compared with 0 hour treatment group.

Mentions: Decreased MMP is an important characteristic of early apoptosis. To investigate whether mitochondria were involved in ICTS-induced apoptosis, we measured MMP in MCF-7 cells by flow cytometry using JC-1 staining. As shown in Figure 4, MCF-7 cells treated with 20 µM of ICTS showed a time-dependent loss of MMP. As expected, the positive control treatment CCCP (10 µM) resulted in a significant decrease in MMP.


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)

Effect of isocryptotanshinone (ICTS) on mitochondrial membrane potential (MMP). MCF-7 cells were treated with 20 µM ICTS for 0, 2, 4, 8, and 20 hours, the MMP was determined by flow cytometer using JC-1 staining. Cells treated with 10 µM carbonyl cyanide 3-chlorophenylhydrazone (CCCP) for 20 minutes were used as the positive control.*p<0.05; †p<0.01 compared with 0 hour treatment group.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 4: Effect of isocryptotanshinone (ICTS) on mitochondrial membrane potential (MMP). MCF-7 cells were treated with 20 µM ICTS for 0, 2, 4, 8, and 20 hours, the MMP was determined by flow cytometer using JC-1 staining. Cells treated with 10 µM carbonyl cyanide 3-chlorophenylhydrazone (CCCP) for 20 minutes were used as the positive control.*p<0.05; †p<0.01 compared with 0 hour treatment group.
Mentions: Decreased MMP is an important characteristic of early apoptosis. To investigate whether mitochondria were involved in ICTS-induced apoptosis, we measured MMP in MCF-7 cells by flow cytometry using JC-1 staining. As shown in Figure 4, MCF-7 cells treated with 20 µM of ICTS showed a time-dependent loss of MMP. As expected, the positive control treatment CCCP (10 µM) resulted in a significant decrease in MMP.

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