Metformin Synergizes with PD-L1 Monoclonal Antibody Enhancing Tumor Immune Response in Treating Non-Small Cell Lung Cancer and Its Molecular Mechanism Investigation

Abstract

Despite non-small cell lung cancer (NSCLC) treatment is proved to be effective using PD-L1 monoclonal antibody (PD-L1 MAb), it is commonly seen in immune-related adverse events reported. We aimed to explore metformin synergized with PD-L1 MAb in treating NSCLC and its potential molecular mechanism. In mice, the transplantable lung cancer models were established and a co-culture system of CD8+T cells and LLC cells was constructed. The anti-tumor effect was assessed by xenograft tumor growth, proliferation signal Ki67 expression, and MTT assays. Immunohistochemistry and western blot assays were also conducted to determine tumor immune response as well as mechanism investigation. The results indicated that tumor volume and cell proliferation were markedly inhibited following metformin synergized with PD-L1 MAb which was more effective than either single metformin or PD-L1 MAb. The cytokines TNF-α, IL-2, and IFN-γ secretion in CD8+ T cells was significantly increased, and the immune response was enhanced by metformin synergized with PD-L1 MAb. Further, the WB results implied that metformin synergized with PD-L1 MAb could activate the AMPK pathway and inhibit mTOR. AMPK inhibitor (Compound C) was added, and the results showed that the anti-tumor effect was reduced in metformin + PD-L1 MAb + CC than in metformin + PD-L1 MAb which indicates the metformin synergized with PD-L1 MAb efficacy was AMPK pathway dependent. In conclusion, metformin synergized with PD-L1 MAb has better efficacy against NSCLC than metformin or PD-L1 MAb alone in an AMPK-dependent way and facilitates increasing CD8+ T cell infiltration and enhancing tumor immune response.