Evaluation of the effect of conditioned media of mesenchymal stem cells treated with ascorbic acid on viability and proliferative behavior of breast cancer cell
Evaluation of the effect of conditioned media of mesenchymal stem cells treated with ascorbic acid on viability and proliferative behavior of breast cancer cell
Fattane Sam Daliri,1Narges Toolabi ,2Mahmood Talkhabi,3,*Farnoosh Attari,4Mousa Kehtari,5
1. Department of Animal Sciences and Biotechnology, Faculty of Life Sciences and Biotechnology, Shahid Beheshti University, Tehran, Iran 2. Department of Animal Sciences and Biotechnology, Faculty of Life Sciences and Biotechnology, Shahid Beheshti University, Tehran, Iran 3. Department of Animal Sciences and Biotechnology, Faculty of Life Sciences and Biotechnology, Shahid Beheshti University, Tehran, Iran 4. Department of Animal Biology, School of Biology, College of Science, University of Tehran, Tehran, Iran 5. Department of Stem Cell Biology, Stem Cell Technology Research Center, Tehran, Iran
Introduction: Breast cancer is one of the most common types of cancer in women. This cancer is a complex of specific malignancies in the mammary glands. Mental problems caused by surgeries, as well as death due to the spread of the disease, are factors that endanger women's health. Nowadays researchers are looking for new method of cancer treatment. The use of stem cells is one of the applied approaches in field of cancer. Among the types of stem cell, mesenchymal stem cells due to their ability to interact with the microenvironments of cancer cells are consider as an appropriate option. conditioned media of mesenchymal stem cells and their secretive factors affect cancer cells. However, the results of application of these cells in the treatment of cancer is controversial. One of the challenges in using stem cells is the limitations on the proliferation and differentiation Potential of these cells for clinical applications. One of the factors affecting stem cells is ascorbic acid. Ascorbic acid is an effective antioxidant on the proliferation and differentiation of mesenchymal stem cells, which also affects the function of cancer cells.
Methods: In this study, investigated the effect of conditioned media of human Adipose-derived Mesenchymal Stem Cells treated with ascorbic acid on proliferative behavior of breast cancer cells. Initially, human Adipose-derived mesenchymal stem cells were constructed from the stem cell center in the first passage, and breast cancer cells (cell line 4T1) in the fifth passage were prepared from the University of Tehran and cultured for use. In order to obtain the desired concentration of ascorbic acid, by reviewing the articles and performing MTT test for 4T1 cells, considering the cell viability above 50% of 4T1 cells, a concentration of 1 mM was selected. Ascorbic acid was treated to mesenchymal cells at a concentration of 1 mM and the resulting conditioned media was collected. The obtained conditioned media was added to 4T1 cells in different groups. In this study, 4 groups were considered and 4T1 cells were treated with these groups for 24 hours and examined. Groups included 4T1 cells treated with conditioned media of mesenchymal stem cells without FBS, without ascorbic acid, the second group, 4T1 cells treated with conditioned media without FBS, contains ascorbic acid with a concentration of 1 mM, the third group, 4T1 cells treated with conditioned media containing 5% FBS, ascorbic acid free and in the fourth group, 4T1 cells treated with conditioned media containing 5% FBS and 1 mM ascorbic acid were determined. Evaluations included cell morphology, apoptosis and cell cycle by Flow cytometry technique.
Results: The results of the effect of conditioned media of mesenchymal stem cells on the morphology of 4T1 cells showed that in groups with ascorbic acid, there were more changes in cells morphology and cell proliferation was reduced. In studies performed by flow cytometry, apoptosis and cell cycle were evaluated. According to the results of 4T1 cell apoptosis, the highest percentage of cells in the apoptotic stage belonged to the group without FBS and had ascorbic acid. Cell cycle analysis showed that, the group in which 4T1 cells were treated with FBS conditioned media and without ascorbic acid, formed the highest percentage of S-phase cells in the cell cycle.
Conclusion: The results of the study showed that the conditioned media of mesenchymal stem cells treated with ascorbic acid, reduces the proliferation of breast cancer cells, therefore, can be effective in controlling breast cancer.