Pterostilbene, a natural stilbene derivative, has garnered significant attention for its antioxidant, anti-inflammatory, and potential anticancer properties. This study aimed to investigate the therapeutic and molecular effects of blackberry derived pterostilbene on HeLa cervical cancer cell line. The present study was conducted in two phases to explore both the phytochemical potential of different extracts of Blackberries and therapeutic potential of isolated natural Pterostilbene on HeLa cell line. Initially, four blackberry extracts (BBM, BBW, BBH, and BBE) were evaluated for their total phenolic & flavonoid content (TPC and TFC) and antioxidant activity. Among them, BBW exhibited the highest phenolic concentration and ABTS radical scavenging activity, indicating its strong antioxidant potential. In the second phase, Pterostilbene was isolated from blackberry and tested for its cytotoxic effects on cancer cell line. Treatment with pterostilbene led to significant inhibition of cell viability, suggesting strong anticancer activity. Importantly, gene expression analysis revealed a marked upregulation of M-CSF and no effect on GM-CSF; these findings highlight the dual importance of natural plant extracts for antioxidant support and pterostilbene as a promising candidate for targeted anticancer therapy through cytotoxic effect on cancer cell line and modulation of immune-related cytokine signaling.