1. 2. Chemistry Department, Faculty of Science, Cairo University.
Anti-proliferative activity of Natural Antitumor Agents (curcumin) against Human Liver and Breast Cancers (In Vitro study)
Samah A  Loutfy1, Nour Tawfik Abdel-Ghani2, Shaimaa Nazir Galal1 ,  Mostafa H. Elberry1, Enas Radwan3, El-Chaimaa B. Mohamed1 , Serag El din I El Behairi4, and Khaled Yehia Farroh5
 1Virology and Immunology Unit, Cancer Biology Department, National Cancer Institute, Cairo University, Egypt.
2. Chemistry Department, Faculty of Science, Cairo University.
3Immunology and Laboratory Bone Marrow transplantation unit, Clinical Pathology Dept, National Cancer Institute, Cairo University.
,4 Egyptian Organization for biological products and vaccines, Agouza, Giza,.
5Nanotechnology and Advanced Materials Central lab, Agricultural Research Center, Egypt.
Human liver and breast cancers are the most common cancer diseases among Egyptian population. Anti-cancer drugs exhibit limited efficacy, associated with several adverse effects. Therefore, safer and more effective chemoprevention for cancer is still required. Curcumin, (Curcumin longa), showed to have anti-inflammatory and/or anti-cancer activity. Our aim is in vitro extensively evaluating the mechanism of apoptotic effects of curcumin against human liver and breast cancers.
Background
Figure 2. EB-stained Gel Electrophoresis of DNA Extraction from Untreated and Treated (a) MCF-7 cell Line with different concentrations of Curcumin after 24 h Lane 1(Ladder 100 bp), Lane 2 untreated MCF-7 (cell control), Lane 3 (treated MCF-7 with 100 μM 0f Curcumin) and Lane4 (treated MCF-7 with 60 µM of Curcumin), Lane 5 (treated MCF-7 with 40µM of Curcumin), Lane 6 (treated MCF-7 with 30 µM of Curcumin), Lane 7 (treated MCF-7 with 12µM of Curcumin) (b) Huh-7 cell Line with different concentrations of Curcumin after 24 h , Lane1 (Ladder 100 bp), Lane 2 untreated Huh-7 (cell control), Lane 3 (treated Huh-7 with 80 µM 0f Curcumin) and Lane4 (treated Huh-7 with 50 µM of Curcumin), Lane 5 (treated Huh-7 with 40µM of Curcumin), Lane 6 (treated Huh-7 with 30 µM of Curcumin) Lane 7 (treated Huh-7 with 12 µM of Curcumin) (c) WISH cell Line with different concentrations of Curcumin after 48h , Lane1 (Ladder 100bp), Lane 2 untreated WISH (cell control), Lane 3 (treated WISH with 50µM 0f Curcumin) and Lane4 (treated WISH with 30 µM of Curcumin).
Curcumin was screened for its cytotoxic effect on Huh7, MCF-7 and Wish as in vitro models of human liver cancer, human breast cancer, and human normal fibroblast cells respectively. DNA fragmentation assays and flow cytometric analysis were performed to evaluate the apoptotic effect on a cellularand molecular levels. Some apoptotic genes expression were determined on a transcriptional level.
Methods
Table1 : Flow cytometric analysis of IC50 concentration of Curcumin on MCF-7, Huh7 and WISH
Curcumin was prepared at a final concentration of 100 µM in DMSO, and IC50 was found to be 41µM, 58µM and 50µM in Huh7, MCF-7 and Wish respectively. MTT results (figure 1) were confirmed by cellular DNA fragmentation (figure 2) which showed lower concentration of cellular DNA after treatment of cells with IC50 compared to untreated cells. Treatment of cells with IC50 generated an increase in the cell population in S phase compared to untreated cells as revealed by flow cytometric analysis (table 1). Apoptotic genes expression indicated more expression of mRNA of Bax gene compared to untreated cells.(data not shown)
Results
Sample
G0-G1 S
G2-M
MCF-7 Control
67.4%
17.6%
14.9%
MCF-7 cells treated with curcumin 60µM
57%
17.8%
24.9%
Huh-7 Control
81.9%
12.0%
Curcumin treated Huh-7 cells with 50µM
93.5%
5.81%
WISH Control
78.6%
11.5%
6.31%
Curcumin treated WISH cells with 50µM
82.3%
7.18%
Figure 1. Effect of different concentrations of Curcumin (100 µM-6.25 µM) on cell proliferation. (a) on Huh7, (b) on MCF-) and (C ) on Wish) cells respectively, as measured by MTT colorimetric assay.
Curcumin is effectively inhibited proliferation of human breast and liver cancer cells. Special technological application will be adopted to improve its bioavailability before its application as an anticancerous treatment in vivo model. 
Conclusion