Study On The CD₄⁺CD₂₅⁺ T Regulatory Cells In Patients With Hepatocellular Carcinoma

Objective: To study the corporation and significance of Tregs cells and foxp3 as well as CCL22 in peripheral blood mononuclear cells and tumor tissues in patients with HCC.Methods: One hundred patients with HCC and twenty healthy subjects were enrolled in this study. CD3⁺,CD4⁺,CD8⁺,NK cell and CD₄⁺CD₂₅⁺ regulatory T cell in peripheral blood mononuclear cells and liver cancer tissues were analyzed by flow cytometry . The mRNA expression of Foxp3 which is the specific marker of Tregs cells and CCL22 was detected by Reverse-transcriptase polymerase chain reaction (RT-PCR), Six healthy volunteers as control. The levels of TGF-βand IL-10 in the same blood sample were detected by ELISA, twenty healthy volunteers were control. The expression of CCL22 in live cancer tissues, adjacent liver tissues and twelve normal liver tissues was tested by IHC.Results: 1.The percentage of the CD3⁺,CD4⁺, CD8⁺, CD4⁺/ CD8⁺and NK cells in peripheral blood of patients with HCC was 57.32±4.69%,33.35±7.01%,23.79±6.30%,1.43±0.47,11.91±2.54%, respectively. And they in control group is 62.95±0.88%,35.85±2.88%,22.03±2.19%, 1.65±0.23,19.42%±1.76%. There was statistical significance between patients with HCC and controls (P<0.05).2.Tregs in peripheral blood in HCC patients was 17.63±4.11%, and the control was7.68±1.32%, which indicated that Tregs in patients with HCC was significantly higher than that in controls (p=0.000).3. Tregs in liver cancer tissues was 5.24±3.83%, and the adjacent liver tissue was 0.73±0.90%, which indicated that Tregs in cancer tissues was significantly higher than that in adjacent liver tissue (p=0.000).4.The level of the TGF-βand IL-10 in peripheral blood in patients with HCC was 161.39±19.09pg/ml and173.48±20.55pg/ml, respectively. And that in health control was67.55±10.25pg/ml and 32.45±7.32pg/ml. There was statistical significance between patients with HCC and controls (P=0.005 and P=0.001).5.The expression of FOXP3 mRNA in peripheral blood of Patients with HCC was higher than that in controls (P<0.01). There was also significantly difference between the expression of CCL22 mRNA in peripheral blood of Patients with HCC(0.31±0.09) and controls (0.15±0.03) (P<0.01).The expression of FOXP3 mRNA in liver cancer tissues (1.28±0.26) was higher than that in adjacent liver tissues (0.48±0.16) (P<0.01)6.The expression of CCL22 in tumor tissues ,adjacent tumor tissues and normal liver tissues was 0.13±0.06,0.09±0.05 and 0.06±0.05,respectively. There was no significantly difference(P=0.155) between adjacent tumor tissues and normal liver tissues.Conclusions: 1 .Cellular immunnity was significantly lower in HCC patients.2.The corporation of Tregs in peripheral blood in HCC patients was significantly higher than that in controls, this result indicated that Tregs might be associated with the low cellular immune function in patients with HCC.3.The levels of the TGF-βand IL-10 in peripheral blood of patients with HCC was significantly higher than that in controls, Tregs might play an important role by TGF-βand IL-10.4.The expression of FOXP3 mRNA in peripheral blood with HCC and in liver cancer tissues was up-regulation, the evidence proved that CD4⁺CD25⁺ might be Tregs.5.The corporation of Tregs in tumor tissues was significantly higher than adjacent tumor tissue, this consequance indicated that Tregs might be associated with immunologic derangement in local microenvironment.6.The over-expression of CCL22 in tumor tissues and adjacent tumor tissue, which was associated with Tregs increasing in patients with HCC.