Journal: Mediators of Inflammation

Article Title: Ze-Qi-Tang Formula Induces Granulocytic Myeloid-Derived Suppressor Cell Apoptosis via STAT3/S100A9/Bcl-2/Caspase-3 Signaling to Prolong the Survival of Mice with Orthotopic Lung Cancer

doi: 10.1155/2021/8856326

Figure Lengend Snippet: ZQT selectively induced G-MDSC apoptosis by activating the STAT3/S100A9/Bcl-2/caspase-3 signaling pathway. (a) MDSCs from the tumor tissue of tumor-bearing mice treated with normal saline, ZQT, STAT3 inhibitor (S3I-201), or ZQT combined with S3I-201 were isolated using the MojoSort Mouse Isolation Kit. The protein expression of Ki67, STAT1, p-STAT1, STAT3, p-STAT3, S100A9, Arg-1, Bcl-2, Bax, cl-PARP, cyt c, and cl-caspase-3 in MDSCs from both normal saline and ZQT-treated mice was confirmed by Western blotting. (b) Lung tumor tissues from normal saline or ZQT-treated tumor-bearing mice were stained using S100A9, Ly6G, Ly6C, CD4, and CD8 antibodies for IHC assay (scale bar: 50 μ M). (c) T-lymphocyte proliferation assay was used to show dose-dependent suppression of T cell proliferation by G-MDSCs isolated from tumor tissue of tumor-bearing mice treated with normal saline or ZQT. CD3 + T cells isolated from tumor stained with 5 μ M CFSE were incubated with G-MDSCs for 72 h. Meanwhile, CD3 and CD28 antibodies were added for CD3 + T cell stimulation. Flow cytometry was used to quantify 72 h CFSE dilution. (d, e) G-MDSCs from tumor tissue of tumor-bearing mice treated with normal saline or ZQT were isolated using flow cytometry. The purity (CD11b + Ly6G + ) was >90% as assessed, and the protein expression of STAT3 and cl-caspase-3 was determined by IF (scale bar: 20 μ M). n = 5. Data are expressed as the mean ± SD. n/s: nonstatistical significance. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001.

Article Snippet: In another experiment, 20 model mice were intragastrically administered with a daily dose of 200 μ L Ze-Qi-Tang, intraperitoneally injected with 5 mg/mL STAT3 inhibitor (S3I-201, CSNpharm), separately or combined, 3 times a week for 4 weeks.

Techniques: Saline, Isolation, Expressing, Western Blot, Staining, Lymphocyte Proliferation Assay, Incubation, Cell Stimulation, Flow Cytometry

Journal: Mediators of Inflammation

Article Title: Ze-Qi-Tang Formula Induces Granulocytic Myeloid-Derived Suppressor Cell Apoptosis via STAT3/S100A9/Bcl-2/Caspase-3 Signaling to Prolong the Survival of Mice with Orthotopic Lung Cancer

doi: 10.1155/2021/8856326

Figure Lengend Snippet: ZQT induced apoptosis in G-MDSCs in TME and inhibited its immunosuppressive activity by activating the STAT3/S100A9/Bcl-2/caspase-3 signaling pathway. Mice were treated with anti-Ly6G neutralizing antibody or isotype control by intraperitoneal injection for 14 days before surgery, after which model mice were given maintenance treatment every other day. (a, b) The bioluminescence images of model mice on day 7 and day 28 after surgery. (c) Survival curves. (d, e) The percentage of G-MDSCs, CD3 + T cells, CD4 + T cells, and CD8 + T cells in tumor tissue were analyzed by flow cytometry. (f, g) CD3 + T cells isolated from the tumor tissue of tumor-bearing C57BL/6 mice in different groups were cocultured with LLC cells for 4 h before they were incubated with CD3, CD8, and CD107 α antibodies. Events shown were finally gated on CD3 and CD8. (h) ELISA analysis for the expression of IFN- γ in cells from tumor tissue. (i) RT-qPCR analysis of Arg-1, iNOS, and S100A9 gene expression and flow cytometry for ROS production. (j) IHC staining for Ly6G and CD8 in tumor tissue (scale bar: 50 μ M). n = 5. Data are expressed as the mean ± SD. n/s: nonstatistical significance. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001.

Article Snippet: In another experiment, 20 model mice were intragastrically administered with a daily dose of 200 μ L Ze-Qi-Tang, intraperitoneally injected with 5 mg/mL STAT3 inhibitor (S3I-201, CSNpharm), separately or combined, 3 times a week for 4 weeks.

Techniques: Activity Assay, Control, Injection, Flow Cytometry, Isolation, Incubation, Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR, Gene Expression, Immunohistochemistry

Journal: Mediators of Inflammation

Article Title: Ze-Qi-Tang Formula Induces Granulocytic Myeloid-Derived Suppressor Cell Apoptosis via STAT3/S100A9/Bcl-2/Caspase-3 Signaling to Prolong the Survival of Mice with Orthotopic Lung Cancer

doi: 10.1155/2021/8856326

Figure Lengend Snippet: The immune regulatory activity of TCM formula ZQT in TME. ZQT induces the apoptosis of G-MDSCs via the STAT3/S100A9/Bcl-2/caspase-3 signaling pathway, resulting in a significant decrease in G-MDSCs and their immunosuppressive activity, consequently promoting the infiltration and killing activity of CD8 + T cells, leading to the inhibition of the proliferation of tumor cells.

Article Snippet: In another experiment, 20 model mice were intragastrically administered with a daily dose of 200 μ L Ze-Qi-Tang, intraperitoneally injected with 5 mg/mL STAT3 inhibitor (S3I-201, CSNpharm), separately or combined, 3 times a week for 4 weeks.

Techniques: Activity Assay, Inhibition