Noscapinoids, synthetic derivatives of noscapine, have been identified as potential anticancer drugs in cervical cancer. This study aimed to understand the mechanism of action of noscapinoids by comparing the gene expression profiles of treated and untreated Hela cells using RNA-seq. A total of 4926 differentially expressed genes (DEGs) were identified, including BAG2, BCL2L11, HSPA4, CASP14, CASP12, CASP8, CASP10, CASP1, and CFLAR. Pathway analysis revealed 20 enriched pathways from up and downregulated DEGs, including endoproteases, cysteine-type endopeptidase activity, TNF, IL-17 signaling pathway, Mitophagy, and Shigellosis. A detailed investigation into DEGs using annotations for cellular components, molecular functions, and biological processes identified different 20 regulating activities from both up and downregulating DEGs. To determine the 50 most functional genes from each group, a protein-protein interaction network was built. These genes were then subjected to further testing by reverse transcription polymerase chain reaction. It was discovered that the downregulated genes that were the focus of the investigation were related to tumor cell invasion, migration and motility, and cell survival and proliferation. The study suggests that these DEGs could be targeted for therapeutic purposes and may serve as potential biomarkers for developing novel therapeutic approaches in cancer treatment. The research sheds light on the mechanism of action of noscapinoids and highlights their potential as a new class of anticancer drugs.