ABSTRACT
Objective To investigate the effect of resveratrol (RSV) on epithelial-mesenchymal transition of MDA-MB-231 cells by down-regulating POLD1 expression. Methods CCK-8 was used to detect the effect of RSV on the activity of MDA-MB-231 cells. POLD1-OE and POLD1-NC cell lines were constructed by transfecting MDA-MB-231 cells with recombinant lentivirus. Western blot was used to detect the expression of POLD1, E-cadherin, N-cadherin and Vimentin after RSV treatment. Transwell invasion experiment and the scratch test were used to detect the cells invasion and migration abilities of each experimental group. Results RSV could significantly inhibit the survival of MDA-MB-231 cells, reduce the expression of POLD1, N-cadherin and Vimentin, increase the expression of E-cadherin, and inhibit the abilities of cell invasion and migration. Increasing the POLD1 expression could reduce the above-mentioned biological effects of RSV on MDA-MB-231 cells. Conclusion RSV could significantly inhibit the viability and EMT of MDA-MB-231 cells by down-regulating the expression of POLD1.
ABSTRACT
BACKGROUND: Studies have shown that adipose-derived stem cells have pluripotent differentiation potential, but only 30%-40% of cells can differentiate into mature adipocytes with low adipogenic differentiation potential. Therefore, how to improve the adipogenic differentiation ability of adipose-derived stem cells is a key problem to be solved in the process of soft tissue regeneration. OBJECTIVE: To observe the relationship between the surface marker CD54 of rabbit adipose-derived stem cells and their adipogenic capacity, and to explore the adipogenic differentiation of CD54+/CD54- adipose-derived stem cells underthe same induction. METHODS: We successfully isolated and cultured the adipose-derived stem cells from inguinal subcutaneous fat pads (3 ml) of New Zealand white rabbits, aged 8-12 weeks, which were induced into multi-differentiation and used to detectsurface markers. We sorted the passage 3 adipose-derived stem cells by immunomagnetic beads and divided into two categories including CD54+ and CD54- adipose-derived stem cells. After 14 days of adipogenic induction, the cells in the two groups were subjected to oil red O staining and were compared by detecting the density of mature adipocytes and lipid droplet contenT.RESULTS AND CONCLUSION: The cultured adipose-derived stem cells possessed the characteristics of mesenchymal stem cells that could differentiate into mature adipocytes, osteoblasts and chondrocytes, with CD29, CD44, CD49d, CD54, CD73, CD90 and CD105 positive expression while CD31, CD34 and CD45 negative expression. Fourteen days after adipogenic induction, the density of mature adipocytes and the intracellular lipid droplet content in the CD54+ group were significantly higher than those in the CD54- group (P < 0.05). We also found that the mRNA expressions of PPARγ,ADD1, C/EBPα related to adipogenic differentiation in the CD54+ group were significantly higher than those in the CD54- group (P < 0.05). Taken together, CD54+ adipose-derived stem cells have excellent adipogenic differentiation capacity.
ABSTRACT
BACKGROUND:Various factors can affect the osteogenic differentiation of human adipose-derived stem cel s, and the osteoinductive factor of traditional Chinese medicine is very important for the research of human adipose-derived stem cel s. OBJECTIVE:To investagate the effects of ginsenoside Rb1 on the proliferation and osteogenic differentiation of human adipose-derived stem cel s in vitro. METHODS:The human adipose-derived stem cel s were isolated and cultured in vitro. After passaqed to the third generation, human adipose-derived stem cel s at 2×103/wel were incubated in a 96-wel plate, and treated with 200μL of 0.5, 1.0, 2.0, 4.0,6.0μmol/L ginsenoside Rb1 medium. The human adipose-derived stem cel s in the control group were treated with an equal volume of Dulbecco’s modified Eagle medium. Growth curves were examined by 2,3-bis-(2-methoxy-4-nitro-5-sulphenyl)-(2H)-tetrazolium-5-carboxanilide T colorimetric assay. Alkaline phosphatase activity and osteocalcin content were detected by alkaline phosphatase kit and radio-immunity method, respectively. Calcified nodules were observed using alizarin red O staining. RESULTS AND CONCLUSION:The proliferation viability of human adipose-derived stem cel s was significantly increased after cultured with 0.5μmol/L ginsenoside Rb1. With the increasing of the concentration of ginsenoside Rb1, the mitogenic activity of the cel s was decreased. The 6.0μmol/L ginsenoside Rb1 showed a depressant effect on proliferation. Ginsenoside Rb1 could promote alkaline phosphatase activity and osteocalcin expression in human adipose-derived stem cel s and showed a dose-dependent manner. Calcified nodule formation induced by 4.6 and 6.0μmol/L ginsenoside Rb1 were better when compared with 0.5, 1.0 and 2.0μmol/L ginsenoside Rb1. Ginsenoside Rb1 can promote the proliferation of human adipose-derived stem cel s cultured in vitro in a certain concentration, and in the high concentration, the ginsenoside Rb1 can promote the osteogenic differentiation of human adipose-derived stem cel s. So ginsenoside Rb1 can be used as an osteoinductive factor.