Global lung cancer rates are rising, prompting the need for effective treatments. The World Health Organization reports that smoking-related lung cancer is the primary cause of cancer-related deaths worldwide. This study focuses on in vitro analysis of cisplatin for lung cancer treatment, examining the role of STAT3, an essential protein in cell signalling and gene expression. The aim is to gain insights into cisplatin's efficacy, optimal dosage, resistance mechanisms, and potential combination strategies to improve lung cancer treatment protocols and patient outcomes. A549 cells (non-small cell lung cancer) were treated with cisplatin in a 15% fatal bovine serum at 37°C with 5% CO2. Cell viability was assessed using the MTT assay. RNA was isolated using the TRIzol method, and cDNA conversion was performed through thermo-cycling. Gene expression analysis was conducted using qRT-PCR with validated primers and SYBR Premix Ex Taq II. Statistical analysis was carried out using SPSS, and results were presented as mean ± SD. Clinical studies support the efficacy of cisplatin and STAT3 treatment in lung cancer, showing improved survival and reduced metastasis with cisplatin-based chemotherapy. Our experimental study aligns with these findings, highlighting cisplatin's potential in inhibiting lung cancer cell proliferation and migration. As a result of cisplatin treatment, lung cancer cell proliferation and migration were found to be significantly decreased in our study. These results align with other studies that have demonstrated the blocking effects of cisplatin and STAT3 on lung cancer cell proliferation and motility.