TGF-B1-over-expressed adipose stem cells-derived secretome exhibits CD44 suppressor and anti-cancer properties via antagonistic effects against SMAD4 in breast cancer cells.
Hasan Salkına, Arzu Yayb, Nur Seda Gökdemirc, Zeynep Burçin Gönenc, Saim Özdamard, Birkan Yakanb
- aDepartment of Medical Services and Techniques, Program of Pathology Laboratory Techniques, Vocational School, Beykent University Istanbul, Turkey
- bDepartment of Histology and Embryology, Faculty of Medicine, Erciyes University Kayseri, Turkey
- cGenome and Stem Cell Center, Erciyes University Kayseri, Turkey
- dDepartment of Histology and Embryology, Faculty of Medicine, Pamukkale University Kayseri, Turkey
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Abstract
OBJECTIVES: This study aimed to investigate the effect of TGF-B1-transfected adipose-derived mesenchymal stem cell (AD-MSC) conditional medium (TGF-B1-CM) on CD44 expression and biological activities in MCF-7 and MDA-MB-231 cells. METHODS: In the study, the experimental groups were created as a standard medium, AD-MSC-CM, TGF-B1-CM, and TGF-B1 recombinant protein. The medium and proteins specified in these groups were applied to MCF-7 and MDA-MB-231 cells separately at 24, 48 and 72 hours. Western blot and immunofluorescent staining were performed with antibodies suitable for CD44 and canonical smad signaling pathway analyses between groups. Cellular proliferation in MCF-7 and MDA-MB-231 cells was measured by MTT. Biological activity analyses such as apoptosis, cell cycle, proliferation, DNA damage, and membrane depolarization between groups were tested on the Muse Cell Analyzer using appropriate kits. Cellular migration between groups was determined by showing cells that migrated to the scar area with in vitro scar formation. Statistics were performed with GraphPad Prism 8.02 software. RESULTS: It was determined that TGF-B1-CM activates the smad signaling pathway in MCF-7 and MDA-MB-231 cells. TGF-B1-CM increased pSMAD2/3 expression and decreased SMAD4 expression in breast cancer cells. A decrease in CD44 expression was found at points of increase in pSMAD2/3 expression. Decreased expression of SMAD4 in breast cancer cells with TGF-B1-CM was associated with decreased expression of CD44. In MCF-7 and MDA-MB-231 cells, TGF-B1-CM was found to increase apoptosis, decrease proliferation, disrupt membrane depolarization, and arrest cells at G0/G1 stage. TGF-B1-CM suppressed MCF-7 and MDA-MB-231 migrations. CONCLUSION: .
TL;DRIt was determined that TGF-B1-CM activates the smad signaling pathway in MCF-7 and MDA-MB-231 cells, and SMAD4-targeted therapeutic strategies may be considered to suppress CD44 expression in breast cancer cells. (Semantic Scholar)