| Objective:To investigate the inhibitory effect of Human Liver Stem Cells(HLSCs)on Ovarian Cancer(OC).Methods:1.In vitro experiment:SKOV-3 cells were divided into RPMI-1640(Control group),HLSCs each concentration of conditioned medium group(25%HLSCs-CM group,50%HLSCs-CM group,75%HLSCs-CM group),and after 48h of group intervention,CCK-8 was measured to detect the inhibition rate of proliferation of SKOV-3 by different proportional concentrations of HLSCs-CM.2.In vivo experiments:primary ovarian cancer cells were isolated from fresh surgical specimens and cultured with microcarrier 6 for 24 h to reach saturation;10 female C57BL/6 mice were randomly divided into two groups,HLSCs intervention group and PBS control group,respectively.200μl HLSCs(containing 4×10~7HLSCs)were injected into the right axilla of the HLSCs intervention group 12 h before modeling,and 200μl PBS was injected into the right axilla of each mouse 12h later.In the HLSCs intervention group,200μl of HLSCs(containing 4×10~7HLSCs)were injected into the right axilla of each mouse 12h before modeling,while the PBS control group was injected with an equal volume of 200μl of PBS,and 100μl of human primary ovarian cancer cells-microcarrier 6 mixture(containing 2.0×10~6tumor cells+30μg microcarrier)were injected into the right axilla of each mouse 12h later,and the tumor-forming volume of transplanted tumors in each group was recorded,Bone marrow and spleen were taken at 16 days of feeding,cell suspensions were prepared,and the cell contents of MDSCs(CD11b+,Gr-1+)and DCs(CD11C+)and the CD4+/CD8+ratio in CD3+T cells were detected by flow cytometry in both groups.Results:1.In vitro experiment:after HLSCs-CM intervened SKOV-3 cells at different ratios(25%,50%,75%)for 48h,the CCK8 results showed that compared with the control group,the inhibition of SKOV-3 cells was9.83±1.47%,33.83±3.31%,44.83±1.17%,respectively,and HLSCs conditioned medium had a had a significant inhibitory effect,with a statistically significant difference,p<0.0001,and the inhibition was enhanced with a higher proportion of conditioned medium,with a statistically significant difference,p<0.0001.2.In vivo experiments:under HLSCs intervention,the tumor-forming volume of the intervention group was significantly smaller than that of the control group,and the difference was statistically significant,p<0.0001.After 16 days of modeling,bone marrow,spleen,and peripheral blood cells were taken from mice and detected by flow cytometry,and the expression of CD11b+,Gr-1+in bone marrow was significantly lower in the HLSCs intervention group compared with the PBS control group.The percentage of CD11b+,Gr-1+MDSCs((？)±s):control group:52.07±2.905%;HLSCs intervention group:41.54±2.505%,the percentage of MDSCs in HLSCs intervention group was significantly reduced,the difference was statistically significant,p<0.0001;the spleen of HLSCs intervention group The percentage of CD11c+DCs was significantly higher in the spleen of HLSCs intervention group((？)±s):PBS control group:0.897±0.156%;HLSCs intervention group:1.914±0.192%,the difference was statistically significant,p<0.0001;CD4+/CD8+T cells ratio in peripheral blood of HLSCs intervention group was significantly higher than that of PBS.T-cell ratio was significantly higher in the HLSCs intervention group than in the PBS control group,((？)±s):PBS control group:1.424±0.063;HLSCs intervention group:2.220±0.115,with a statistically significant difference,p<0.0001.Conclusion:HLSCs can inhibit ovarian cancer cell proliferation and reshape the body’s immune system by down-regulating the body’s immunosuppressive cells MDSCs and up-regulating anti-tumor immune cells DCs and CD4+/CD8+T ratio,thus inhibiting the progression of ovarian cancer. |
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