Influence of two indigenous plant extracts on proliferation, migration and adhesion of breast cancer cell lines Munmi Majumder and Rupak Mukhopadhyay Department of Molecular Biology and Biotechnology, Tezpur University, Assam Email:
[email protected];
[email protected] Abstract Breast cancer is currently the leading cause of cancer-related deaths among women globally. Notwithstanding current therapeutic regimen available for treatment of the disease, studies are ongoing to find more safe and potent therapies. Several medicinal plant extracts has been shown to have potential impact against cancer. In our laboratory we study the effects of indigenous plants on breast cancer cells. Metastasis is an important characteristic of cancer cells which can be targeted as a therapeutic strategy. In this study, we report the efficacy of two plant-extracts FEC and REC (code names) on proliferation, migration and adhesion of MCF-7 (low-metastatic potential) and MDA-MB-231 (high metastatic potential) cells. Results from MTT assay revealed that both extracts (in water : ethanol= 50:50) inhibited the proliferation of MCF-7 and MDA-MB-231 cells in a time and dose-dependent manner. To establish the anti-cancer efficacy, we further studied the anti-metastatic property of these extracts. Both extracts significantly inhibited the migration of the cells in wound healing assay after 12 and 24 hours of incubation. Cell adhesion in collagen IV coated wells was reduced by incubation with the plant extracts in a concentration-dependent manner. Studies on molecular mechanism of action of these extracts are ongoing. Our data suggests that REC and FEC can be potent candidates for therapies against breast cancer. Incidences: Breast cancer scenario in India
Inhibition of migration of MCF-7 and MDA-MB-231 cells by FEC and REC
2012
2008
10 Most commonly diagnosed cancers
Anti-proliferative efficacy of FEC and REC on MCF-7 and MDA-MB-231 cells
FEC and REC showed cytotoxicity in a dose dependent as well as time dependent manner in the cell lines MCF-7 and MDA-MB-231. Both the extracts showed better efficacies against MDA-MB-231 cells compared to MCF-7 cells. In case of MCF-7 the % of viable cells at 250µg/ml of FEC is 68% in 24 hour which is reduced to 54% at 48 hour. But in case of MDA-MB-231 cells the % of live cells at the same concentration is 46% at 24 hour and 28% at 48 hour. Treatment with 250µg/ml of REC for 24 hr showed 72% viability of MCF-7 cells which is reduced to 62% at 48 hour whereas the % of live cells at 24 hour treatment in case of MDA-MB-231 is 59% and is reduced to 34% at 48 hour.
Inhibition of adhesion of MCF-7 and MDA-MB-231 cells by FEC and REC After incubation of 2x105 cells/ml in serum free media cells were pretreated with different doses of extracts for 1hr. Cells were plated in 96well collagen IV coated plates and allowed to adhere for 60 min, the medium was gently removed and the wells were washed, and attached cells were assayed using MTT assay. Both the extracts inhibited adhesion in both the cell lines in a dose dependent manner. Treatment with 20ug/ml FEC inhibited adherence by 25% in MCF-7 and 16% in MDA-MB-231 while by REC inhibited adherence by 17% in MCF-7 and 28% in MDA-MB-231.
Wound healing assay was performed for both the cell lines MCF-7 and MDA-MB-231. Confluent monolayer of cells were pre-treated with 1ug/ml concentration of mytomicin for 1hr and then a scratch was made using 10ul pipette tip. The cells were washed and treated with extracts. Pictures were taken at 3 different fields at 0, 24 and 48hrs. FEC has significantly decreased the migration in MDA-MB-231 in a very low concentration of 0.05 µg/ml and 0.1 µg/ml. Similarly REC also significantly decreased the migration at a concentration of 10 µg/ml and 20 µg/ml in both the breast cancer cells .
Conclusion These data suggests that indigenous plant extracts FEC and REC have a strong anti-proliferative activity and also have inhibited adhesion and migration significantly. These extracts can inhibit metastasis of breast cancer as well as can act as a potent candidate against breast cancer.
Future prospects
To investigate the extracts for inhibition of invasion To study the molecular mechanism of anti-metastasis action To study the mechanism of anti-cancer action To separate and identify the active components of the extracts To develop a drug regimen from the active component/s
References Chun, Jaemoo, and Yeong Shik Kim. "Platycodin D inhibits migration, invasion, and growth of MDA-MB-231 human breast cancer cells via suppression of EGFR-mediated Akt and MAPK pathways." Chemico-biological interactions205.3 (2013): 212-221. Wang, Ling, et al. "Flavonoid baicalein suppresses adhesion, migration and invasion of MDA-MB-231 human breast cancer cells." Cancer letters 297.1 (2010): 42-48. Phannasil, Phatchariya, et al. "Pyruvate Carboxylase Is Up-Regulated in Breast Cancer and Essential to Support Growth and Invasion of MDA-MB-231 Cells."PloS one 10.6 (2015): e0129848.Wang, Ling, et al. "Flavonoid baicalein suppresses adhesion, migration and invasion of MDA-MB-231 human breast cancer cells." Cancer letters 297.1 (2010): 42-48.
Acknowledgements The authors gratefully acknowledge the funding received under DBT-Twining project and DBT U-EXCEL project from Department of Biotechnology (DBT) to RM for carrying out the project. MM acknowledges the scholarship from DST-INSPIRE program.
