<p>The incidence and mortality of non-small cell lung cancer (NSCLC) remain high in China. Recently, several studies have shown a complex correlation between microflora and NSCLC. Compared to patients with healthy/benign lung disease, patients with NSCLC have distinct microflora within the tumor, airways as well as intestines. NSCLC can be diagnosed with the assistance of characteristic microflora and circulating microbial DNA. Microflora changes can affect the efficacy of immunotherapy and treatment-related adverse effects during lung cancer therapy. Modulating microflora imbalance through antibiotics/probiotics nebulization, oral probiotic, prebiotics, and dietary modifications can balance microflora and enhance the effectiveness of immunotherapy. Therefore, microbe-specific testing and targeted therapy have the potential to become a new method in the diagnosis and treatment of NSCLC. This review will explore the molecular mechanisms by which microbes influence lung cancer, and summarize the latest research advances in microbial biomarkers and therapeutic approaches for NSCLC.</p>