Paper Title
BIOTECHNOLOGICAL APPLICATIONS IN UNCOVERING THE ANTICANCER POTENTIAL OF POLYGONUM BISTORTAL

Abstract
Polygonum is an important genus in the Polygonaceae family, with 35 species in Iran. P.bistortaL. is one of the medicinal species in this genus, known for its anticancer properties against HT_29 colon cancer cells. Methanolic extracts of the leaves and rhizomes of P.bistortaL. were prepared, and their antioxidant and anticancer effects on HT-29 cells were evaluated using MTT assay, GC-MS analysis for bioactive compound identification. Then, to assess the changes in the expression of apoptosis-related genes, RNA extraction, cDNA synthesis, and Real-time PCR were performed using specific primers for the target genes. Bioactive compounds, including pyrocatechin, 1,2,3-benzenetriol, and β-tocopherol, were identified as important components with anticancer properties that reduce cellviability. Additionally, the methanolic extract of Polygonum was able to down regulate the expression of the BCL2 gene while up regulating the expression of the BAX gene. These changes in gene expression indicate the activation of apoptotic pathways in HT-29 cancer cells.Overall, the methanolic extract of the rhizomes effectively inhibited the growth of HT-29 cancer cells, suggesting its potential as a therapeutic agent.Based on our results, Polygonum bistortaL. Is not only valuable for its anticancer properties but also for a comprehensive analysis of molecular mechanisms and key cancer pathways. By integrating advanced techniques such as GC-MS and Real-Time PCR, this study offers a comprehensive understanding of molecular interactions and the therapeutic potential of this plant in cancer treatment. These findings could play a significant role in the development of natural anticancer drugs based on Polygonum bistortaL. and open new avenues for future research in this field. Keywords - Polygonum bistorta L.; HT-29 Colon Cancer Cells; GC-MS; Anticancer Compounds; Real-Time PCR