Abstract
Background: Melanoma, an aggressive form of skin cancer, has garnered substantial research attention due to its rising incidence and limited treatment efficacy. Tumor cells often become more aggressive post-chemotherapy, evading innate and adaptive immune responses, leading to recurrence and metastasis. Identifying mechanisms of chemotherapy resistance is crucial to reduce mortality rates. Negative immune pathways such as cytotoxic T-lymphocyte-associated protein 4 (CTLA-4), programmed death-ligand 1 (PD-L1), and V-domain Ig suppressor of T cell activation (VISTA) hinder immune responses to cancer. This study aimed to uncover the dynamic alterations in the expression patterns of key immune checkpoints VISTA, PD-L1, and CTLA-4 in melanoma cancer cell line.
Methods: In this study, the A-375 melanoma cancer cell line was cultivated using conventional cell culture techniques. The IC50 (half-maximal inhibitory concentration) and efficacy assessment of chemotherapy drugs, docetaxel and doxorubicin, were determined using the MTT test. After treatment, an analysis of the expression of PD-L1, CTLA-4, and VISTA genes in the A-375 cell line was performed using quantitative reverse transcription-polymerase chain reaction (qRT-PCR). One-way analysis of variance (ANOVA) was used for statistical analysis. All experiments were conducted in triplicate, with significance set at P<0.05. Statistical significance is denoted as follows: *P<0.05, **P<0.01, ***P<0.001, and ****P<0.0001.
Results: Significant upregulation of VISTA expression was observed in both ADO and ADC groups. CTLA-4 expression increased significantly in the ADC group, while no significant change was observed in the ADO group. PD-L1 expression was significantly higher in both ADO and ADC groups compared to the control group.
Conclusion: These compelling findings underscore the profound impact of chemotherapeutic agents on the heightened expression of immune checkpoint genes, shedding light on the potential modulation of crucial pathways associated with immune response regulation.