J Breast Cancer.  2014 Sep;17(3):219-225. 10.4048/jbc.2014.17.3.219.

Antiproliferatory Effects of Crab Shell Extract on Breast Cancer Cell Line (MCF7)

• Rezakhani
• Rashidi Z
• Mirzapur P
• Khazaei M

• KMID: 2286348
• DOI: http://doi.org/10.4048/jbc.2014.17.3.219

Abstract

PURPOSE
Breast cancer is the most common type of cancer in women. Despite various pharmacological developments, the identification of new therapies is still required for treating breast cancer. Crab is often recommended as a traditional medicine for cancer. This study aimed to determine the in vitro effect of a hydroalcoholic crab shell extract on a breast cancer cell line.
METHODS
In this experimental study, MCF7 breast cancer cell line was used. Crab shell was powdered and a hydroalcoholic (70degrees ethanol) extract was prepared. Five concentrations (100, 200, 400, 800, and 1,000 microg/mL) were added to the cells for three periods, 24, 48, and 72 hours. The viability of the cells were evaluated using trypan blue and 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays. Cell apoptosis was determined using the terminal deoxynucleotidyl transferase dUTP nick end labeling method. Nitric oxide (NO) level was assessed using the Griess method. Data were analyzed using analysis of variance, and p<0.05 was considered significant.
RESULTS
Cell viability decreased depending on dose and time, and was significantly different in the groups that were treated with 400, 800, and 1,000 microg/mL doses compared to that in the control group (p<0.001). Increasing the dose significantly increased apoptosis (p<0.001). NO secretion from MCF7 cells significantly decreased in response to different concentrations of the extract in a dose- and time-dependent manner (p<0.050).
CONCLUSION
The crab shell extract inhibited the proliferation of MCF7 cells by increasing apoptosis and decreasing NO production.

Keyword

Apoptosis; Breast neoplasms; Cell survival

MeSH Terms

Apoptosis
Breast Neoplasms*
Cell Line*
Cell Survival
DNA Nucleotidylexotransferase
Female
Humans
MCF-7 Cells
Medicine, Traditional
Nitric Oxide
Trypan Blue
DNA Nucleotidylexotransferase
Nitric Oxide
Trypan Blue

Figure

• 

  Figure 1 (A) MCF7 viability (%) after 24, 48, and 72-hour exposed to different doses of crab shell extract. Significant difference between the experiments and control groups were seen. *p<0.05; †p<0.001. (B) MCF7 cells in control and different doses of the extract at 72 hours by invert microscope (×100).

• 

  Figure 2 MCF7 cell line viability (%) in different crab shell extracts and different culture periods. There is significant difference between the groups. *p<0.05; †p<0.001.

• 

  Figure 3 Nitric oxide (NO) levels in different groups and different culture periods. Significant difference between groups. *p<0.05.

• 

  Figure 4 (A) MCF7 apoptotic index (%) after 72-hour with different amounts of crab shell extract. Significant difference between groups. *p<0.05; †p<0.001. (B) Identification of apoptosis in MCF7 cell line following terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining and counterstaining with propidium iodide after 72 hours of exposure crab shell extract. TUNEL positive cells (apoptotic nuclei) identified by a distinct bright yellow stained chromatin (arrows). C+: The positive control was incubated with ethanol 10% for 10 minutes, all cells are seen as bright yellow. C-: The positive control was incubated with label solution without enzyme solution. No TUNEL positive cells were observed (×200).

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