Study On The Anti-Liver Cancer Of CTL Activated By Dendritic Cell Vaccine Loaded With RNA From CD133+LCSC

Background and Objective:Liver cancer is one of the common malignant tumor in our country,although liver cancer treatment such as surgery,chemotherapy and radiotherapy level in ris ing,but the treatment effect of liver cancer is still not ideal,so it is particularly important to find new effective treatment.Tumor immunotherapy is a new method after traditional tumor therapy and is considered to be one of the most promising treatments for cancer.Dendritic cell(DC)can induce highly effective and specific anti-tumor immune response,and the research on anti-tumor based on dendritic cells has become a research hotspot.In our previous studies,we have successfully prepared dendritic cell vaccine loaded with RNA from Liver cancer stem cells(LCSC).This study used dendritic cell vaccine loaded with RNA from LCSC to activate cytotoxic T lymphocyte(CTL),and investigated the effect of CTL on the killing activity of hepatocellular carcinoma cells in vitro and anti-liver cancer in nude mice.The objective is to provide an experimental basis for the further study of dendritic cell vaccine to treat liver cancer.Methods:First of all,hepatocellular carcinoma cells were segregated from human HCC tissue by the enzyme digestion method,and CD133+hepatocellular carcinoma cells were sorted by flow cytometry,and RNA of cells was extracted,the dendritic cell vaccine loaded with RNA from CD133+hepatocellular carcinoma cells were prepared.Then,the activated CTL were induced by the dendritic cell vaccine loaded with RNA from CD133+hepatocellular carcinoma cells,and the levels of gamma interferon secreted by CTL were detected by ELISA.Finally,the ability of CTL to kill cells in vitro and tumor inhibition in nude mice were detected.Results:Thecontentofgammainterferonin CD133+HCC-RNA-DC-CTL group,CD133-HCC-RNA-DC-CTL group,mDC-CTL group and T lymphocyte group in the supernatant were(926.50±40.32),(892.20±35.57),(880.25±32.21)and(302.47±21.43)pg/ml respectively,there was no statistically s ignificant difference in the content of gamma interferon in the first three groups(P?0.05),the first three groups were all higher than T lymphocyte group,and the differences were statistically significant(P<0.05).In vitro cell damage experiment,when the target cells were CD133+liver cancer stem cells,the killing activity of each group was compared.CD133+HCC-RNA-DC-CTLgroupishighest,followedby CD133-HCC-RNA-DC-CTL group,the mDC-CTL group minimum,there are statistically significant difference between the three groups(P<0.05);When the target cells were CD133-hepatocellular carcinoma cells,the killing activity of each group was compared.CD133-HCC-RNA-DC-CTL group is highest,followed by CD133+HCC-RNA-DC-CTL group,mDC-CTL group minimum,there are statistically s ignificant difference between the three groups(P<0.05).In vivo tumor suppressive experiments,CD133+HCC-RNA-DC-CTL group,CD133-HCC-RNA-DC-CTL group,mDC-CTL group and control group began to become tumor at week 5,week 4,week 2 and week 1 respectively.The tumor volume of CD133+HCC-RNA-DC-CTL group,CD133-HCC-RNA-DC-CTL group,mDC-CTL group and control group were(257.63±36.13)mm~3,(513.40±68.52)mm~3,(1014.52±86.54)mm~3 and(1354.27±84.48)mm~3respectively,The differences between the four groups were statistically significant(P<0.05).Conclusion:CTL activated by the dendritic cell vaccine loaded with RNA from CD133+hepatocellular carcinoma cells can secrete high levels of gamma interferon,it has efficent specific killing activity for liver cancer stem cells and obvious inhibition for the growth of tumor in nude mice,the dendritic cell vaccine loaded with RNA from liver cancer stem cells may be effective vaccine for the treatment of liver cancer.