| In our country,liver cancer is one of the top five diseases which have highest incidence and mortality rates,seriously affecting the quality of life and the life cycle.The best mothods to treat the early liver cancer are surgery and liver transplantation.But to advanced liver cancer,we lack of effective treatment.The regulation of body’s immunity is an auxiliary method of treating advanced liver cancer.It can improve the life quality and prolong the survival time of patients.Cellular immunity mediated by Treg,Th17,Th1 and Th2 cells is an important part of tumor immunity.In cancer patients,the imbalance of Th17/Treg and Th1/Th2 is common.Restoring their balance state is one idea to treat the cancer.Our previous studies demonstrated that recombinant prothymosin a had a good effect on prolonging the survival time of mices bearing cancerous ascites,and had a good antitumor effect in combination with IL-2.However,the effect on Treg,Th17,Th1 and Th2 cells in mices bearing liver cancer is unknown.This experiment medicates the mices in early and advanced stages. We observe the effect on the tumor inhibition rate of the mices bearing liver cancer,and observe if recombinant ProTa can regulate Treg,Th17,Th1 and Th2 cells.36 KM mices were randomly divided into control group,tumor-bearing group and drug intervention group,and a tumor-bearing mice model was established by transplanting H22 cells subcutaneously. By medicating recombinant ProTa through intraperitoneal injection,we observed the tumor inhibition rate and detected the Treg cells percentage by flow cytometry after 7 days and 14 days.We also detected the transcriptional level of specific cytokines of Treg,Th17,Th1 and Th2 cells(Foxp3、RORγt、T-bet、GATA3) by realtime PCR to investigate the underlying mechanism.The results show that the distinguish of tumor weight between tumor-bearing group and drug intervention group is not significant after bearing tumor 7 days.However,after bearing tumor 14 days,the tumor weight of drug intervention group reduces significantly.Detecting the proportion of CD25+Foxp3+Treg cells in CD4- cells by flow cytometry,we observe that the Treg cells proportion of tumor-bearing group is significantly higher than the control group after 14 days tumor-bearing.No matter 14 days intraperitoneal injection of recombinant ProTa or drug intervention from the advanced stage,the proportion of Treg cells declines significantly.The results of Real-time PCR show that the T-bet expression in tumor-bearing group is lower than control group,but the GATA3 expression is higher.It appears that the number of Th2 cells rises in tumor-bearing group.However,after medicating recombinant ProTa,the T-bet expression of drug intervention group rises,and the GATA3 expression declines.Recombinant ProTa can increase the expression of Thl cells and restore the balance of Thl/Th2 cells to inhibit tumor growth.On the other hand,we also find that there is a high expression of RORyt and Foxp3 in mices of tumor-bearing group.After intraperitoneal injection of recombinant ProTa,the expression of RORyt and Foxp3 decreases.In liver cancer-bearing mice model,Thl7 and Treg cells may be jointly involved in promoting the growth of liver cancer cells in tumor-bearing mice,and the anti-tumor effect of recombinant ProTa may be related to the reduction of Th17 and Treg cells expression.In this study,we simulate the drug intervention in early stage and advanced stage by intraperitoneal injection of recombinant ProTa from the first day and the eighth day.The results suggest that recombinant ProTa inhibits tumor growing,and the early and long-term drug intervention benefits the most.The putative mechanism is related to decreasing the proportion of Treg and Th17 cells and restoring the balance of Thl/Th2 cells to inhibit the immune escape of hepatocellular carcinoma cells and increase the anti-tumor effect.And it provides a theoretical basis for further clinical application. |
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