Effects of moxibustion on Treg cells in sarcoma microenvironment

To investigate the therapeutic effect of moxibustion on sarcomas from mesenchymal tissues, which have a low response rate to chemotherapy and radiotherapy.

S180 sarcoma cell line was inoculated in C57BL/6 mice to form transplanted tumor. Moxibustion therapy was directly applied at the transplanted tumor sites, at a distance of 3.0 cm, 10 min per session, till skin temperature reached 45 °C, once a day, for 14 consecutive days of intervention. After the mice were killed, serum was collected and used to detect concentrations of interleukin-10 (IL-10), transforming growth factor-β1 (TGF-β1), IL-4 and interferon-γ (IFN-γ) by Luminex liquid suspension chip. The numbers of Treg⁺ T cells and CD4⁺CD25⁺Forkhead Box P3 (Foxp3)⁺ T cells were detected by flow cytometry. Fluorescence in situ hybridization was used to analyze the changes of CD4, CD8, Foxp3 and TGF-β1 in the tumor microenvironment (TME).

Weight of S180 transplanted tumor in the control group was (2.03 ± 0.54) g, and that in the moxibustion group was (1.27 ± 0.29) g, which was statistically different (P = 0.023). The mean value of Foxp3⁺ T cells in the normal group was 2.01%, which increased to 3.63% after the formation of transplanted tumor, and decreased to 1.48% after moxibustion treatment. The moxibustion group also had reduced numbers of CD4⁺CD25⁺Foxp3⁺ T cells in the spleen of mice with transplanted tumor. The concentrations of IL-10, TGF-β1 and IL-4 decreased in the serum of mice with transplanted tumor, while the concentration of IFN-γ increased. Moxibustion was associated with downregulation in expression of Foxp3, IL-10 and TGF-β1 genes in the transplanted tumor, and increases in the gene expression of CD4⁺ T cells and CD8⁺ T cells in the TME.

Moxibustion may have therapeutic effects on sarcomas by reducing the number of Treg cells in the blood and controlling the infiltration of Treg cells in the TME.