Evaluation of the effect of Dichloroacetate-treated mesenchymal stem cell conditioned media on viability and proliferative behavior of breast cancer cells
Evaluation of the effect of Dichloroacetate-treated mesenchymal stem cell conditioned media on viability and proliferative behavior of breast cancer cells
Narges Toolabi,1Fattane Sam Daliri,2Mahmood Talkhabi,3,*Farnoosh Attari,4Leila taghiyar,5
1. Department of Animal Sciences and Marine Biology, Faculty of Life Sciences and Biotechnology, Shahid Beheshti University, Tehran, Iran 2. Department of Animal Sciences and Marine Biology, Faculty of Life Sciences and Biotechnology, Shahid Beheshti University, Tehran, Iran 3. Department of Animal Sciences and Marine Biology, 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 Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran
Introduction: Cancer is a disease caused by the overgrowth of malignant cells that can spread to other parts of the body. The development of breast cancer is a multi-stage process that begins in the cells of the breast and can be transmitted to distant organs. Drugs that affect cancer metabolism have been studied in recent decades. Many cancer drugs and cancer drug compounds have individual side effects. Dichloroacetate (DCA) has been studied as a new metabolic treatment for various cancers. The effects of stem cells or the conditioned media produced by these cells in the treatment or control of some cancers and tissue repair have already been demonstrated. This study investigates the effect of conditioned media of DCA-treated mesenchymal stem cells on the proliferative behavior of breast cancer cells based on cellular and morphological studies.
Methods: In this study, human adipose-derived mesenchymal stem cells were first prepared in passage 1 and propagated in the laboratory. Then, in passage 3, different doses of DCA were treated and after 24 hours, the viability rate was assessed by MTT assay. Also, 4T1 cell line was prepared in Passage 5 from the University of Tehran. Then, 4T1 cell line in passage 10, different doses of DCA were treated and after 24 hours, the viability rate was assessed by MTT assay. Then a dose of 1 mM was selected to collect the conditioned media. To prepare the conditioned media of human adipose-derived mesenchymal stem cells in passage three with four groups included: group without FBS and without DCA, group without FBS and with 1 mM DCA, group with 5% FBS and without DCA and group with 5% FBS and treated with DCA 1 mM and after 24 hours of conditioned media was collected for further experiments. Then 4T1 cells in 10 passages were treated with concentrations 100% conditioned media of 4 groups. Evaluations cell morphology, apoptosis and cell cycle by Flow cytometry technique were performed.
Results: Study of cell cycle and apoptosis on 4T1 cells treated with 100% conditioned media of group without FBS and with DCA compared to group without DCA, are able to induce S-phase cell cycle and induce apoptosis 4T1 cells. Comparison of the group with 5% FBS and with DCA compared to the group without DCA is able to induce S-phase cell cycle and inhibit apoptosis 4T1 cells. The results of the effect of conditioned media of mesenchymal stem cells on the morphology of 4T1 cells showed that in groups with DCA 1 mM, there were more changes in cells morphology and cell proliferation was increased.
Conclusion: The results showed that the conditioned media treated with low doses of DCA causes proliferation and cessation of apoptosis of 4T1 breast cancer cells.