ABSTRACT
OBJECTIVE@#To study placenta-derived mesenchymal stem cells with HLA-G (Human Leukocyte Antigen, HLA-G) positive expression induce Treg (regulatory T cell, Treg) in vitro.@*METHODS@#placenta-derived mesenchymal stem cells were separated from neonatal placenta; PEGFP - N1 -HLA-G plasmid was transfected in placenta-derived mesenchymal stem cells by liposome transfection.The cells were divided into 3 groups including control group, PEGFP-N1 group and PEGFP-N1-HLA-G group, 5 complex walls in each group. Expression of HLA-G protein was detected by Western Blotting; after identification of cells, healthy human peripheral blood CD4 T lymphocytes were cultured with placenta-derived mesenchymal stem cells with HLA-G positive expression, and the ratio of CD4CD25Foxp3Treg in T lymphocytes was accounted.@*RESULTS@#After transfection of PEGFP-N1-HLA-G, the placenta-derived mesenchymal stem cells can express HLA-G protein significantly, compared with the control group and PEGFP - N1 group (<0.01). After HLA-G positive placenta-derived mesenchymal stem cells and CD4 + T lymphocytes were cultured for 24 h, the ratio of CD4CD25Foxp3Treg in T lymphocytes was (16.41±0.94)%. After HLA - G positive placenta-derived mesenchymal stem cells and CD4 T lymphocytes were cultured for 48 h, the ratio of CD4CD25Foxp3Treg in T lymphocytes was (16.46±0.59)% significantly, compared with the control group and PEGFP - N1 group (<0.01).@*CONCLUSIONS@#Placenta-derived mesenchymal stem cells modified by HLA-G gene can effectively induce CD4CD25Foxp3Treg in vitro.
Subject(s)
Female , Humans , Pregnancy , Forkhead Transcription Factors , HLA-G Antigens , Mesenchymal Stem Cells , Placenta , T-Lymphocytes, RegulatoryABSTRACT
Objective: To evaluate the effects of ropivacaine on cell proliferation and tumor growth of colon carcinoma. Methods: CCK8 assay was performed for cell viability. The colony formation assay was performed for cell proliferation. Cell flow apoptosis and cell cycle were measured by Flow cytometry. Protein levels were calculated by Western blot. Meanwhile,nude mice were inoculated with SW480 colon cells and treated with ropivacaine. Tumor volume and survival rate were examined after treatment. Results: The results of CCK8 showed that the optimum concentrations of ropivacaine were 20, 50 and 100 μg/ ml respectively. Ropivacaine dose-dependently inhibited colony formation (17. 80±0. 51,P<0. 001) and expressions of Ki67 (0. 32±0. 68, P<0. 01) and PCNA(0. 14±0. 24,P<0. 01). Meanwhile,treatment with ropivacaine markedly increased apoptosis (12. 80±1. 24,P< 0. 01) and protein levels of caspase-3(1. 76±1. 43,P <0. 001) and caspase-9 (1. 61±1. 26,P <0. 001) . In addition,ropivacaine notably induced cell cycle arrest (40. 5%,P<0. 01),expression of p53 (1. 16±0. 65,P<0. 01),and down-regulated the expression level of Cyclin A (0. 12±0. 12,P<0. 05) . Furthermore,ropivacaine inhibited tumor growth (1 247. 60±1. 37,P<0. 01),up-regulated survival rate of mice and induced apoptosis of tumor tissue (78. 00 ±1. 45,P <0. 001) in a dose-depended manner. Conclusion: Ropivacaine inhibits cell proliferation and tumor growth of colon cancer.