Background: Non-small cell lung cancer (NSCLC) is often harmful to human health. It is common in people who smoke or smoke passively, and the affected people are older and find it difficult to recover. The main treatment methods are surgery, radiotherapy and chemotherapy. This paper aims to study the effects of different configurations of ginsenoside Rh2 on NSCLC 95D and NCI-H460 cells. To provide clinical value for the treatment of NSCLC. Materials and Methods: CCK-8 was used to determine the proliferation of 95D and NCI-H460 cells with different concentrations of ginsenoside Rh2 (0, 0.05, 0.1 and 0.2 mg/mL), and IC50 was calculated. Cell cycle and apoptosis were detected by immunofluorescence staining and flow cytometry. Results: 20(R)-ginsenoside-Rh2 and 20(S)-ginsenoside-Rh2 inhibited the proliferation and colony formation of NSCLC 95D and NCI-H460 cells induced cell cycle arrest in a time and dose-dependent manner in G1/S and promote apoptosis. Conclusion: Ginsenoside Rh2 induces antitumour activity by inhibiting proliferation and colony formation of 95D and NCI-H460 cells and inducing cell cycle arrest and apoptosis. Comparing the two configurations of ginsenoside Rh2, the effect of 20(R)-ginsenoside-Rh2 is stronger.