J Breast Cancer.  2015 Jun;18(2):112-118. 10.4048/jbc.2015.18.2.112.

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

• Zhang X
• Luo W
• Zhao W
• Lu J
• Chen X

• KMID: 2374525
• DOI: http://doi.org/10.4048/jbc.2015.18.2.112

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.

Keyword

Apoptosis; Breast neoplasms; Isocryptotanshinone; Mitogen-activated protein kinases

MeSH Terms

Apoptosis*
Blotting, Western
Breast Neoplasms*
Cell Cycle
Cell Cycle Checkpoints
Cell Line
DNA Fragmentation
Flow Cytometry
G1 Phase
Hep G2 Cells
Humans
Liver Neoplasms
Lung Neoplasms
MCF-7 Cells*
Membrane Potential, Mitochondrial
Mitogen-Activated Protein Kinases
p38 Mitogen-Activated Protein Kinases
Phosphorylation
Salvia miltiorrhiza
Mitogen-Activated Protein Kinases
p38 Mitogen-Activated Protein Kinases