Thesis_Defense_Presentation_Aizaz khan.pptx

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1. Association Between XRCC1 Variant (rs25487) With Breast Cancer in Affected Population of KPK Presented by: Muhammad Aizaz Khan Department of Biotechnology, University of Swabi June 2025

2. Introduction • Breast cancer: Most common and deadliest cancer in women worldwide. • One in nine Pakistani women may develop it. • XRCC1 gene is critical in base excision repair (BER) of DNA. • rs25487 (Arg399Gln) polymorphism may affect DNA repair function. • No previous research in KPK population.

3. Problem Statement • XRCC1 rs25487 linked to breast cancer risk in various populations. • No study conducted in Khyber Pakhtunkhwa (KPK), Pakistan. • Aim: To investigate its association in local female population.

4. Aims and Objectives Main Aim: • Determine the association of XRCC1 rs25487 with breast cancer in KPK. Objectives: • Assess genotype distribution in patients and controls. • Analyze correlation with breast cancer risk and treatment response.

5. Materials and Methods (Part 1) Step 1: Ethical Approval - Approved by Dept. of Biotechnology, University of Swabi. Step 2: Selection - 100 breast cancer patients (IRNUM), 100 healthy controls. Step 3: Inclusion/Exclusion Criteria - Confirmed cases, no family history in controls. Step 4: DNA Extraction - Phenol–chloroform method.

6. Materials and Methods (Part 2) Step 5: RBC Lysis - Solution A used. Step 6: WBC Lysis - Solution B + SDS used. Step 7: Protein Removal - Phenol:Chloroform:Isoamyl Alcohol (24:1). Step 8: DNA Purification - Chloroform cleaning. Step 9: DNA Precipitation - Isopropanol + Sodium Acetate. Step 10: Washing - 70% ethanol. Step 11: Storage - 20 µL nuclease-free water at –20°C.

7. Materials and Methods (Part 3) Step 12: DNA Confirmation - 1% gel electrophoresis. Step 13: Genotyping - Tetra-primer ARMS-PCR with specific primers. Step 14: Amplicon Sizes - Control: 480bp, G: 247bp, A: 163bp. Step 15: PCR Conditions - Optimized thermal profile. Step 16: PCR Visualization - 2% agarose gel under UV. Step 17: Statistical Analysis - GraphPad Prism, p < 0.05.

8. Results Summary • No significant association found in all genetic models: - Co-dominant: p = 0.537 - Homozygous Dominant: p = 0.689 - Homozygous Recessive: p = 0.774 - Heterozygous Dominant: p = 0.584 - Additive Model: p = 0.829

9. Conclusion • XRCC1 rs25487 does not significantly affect breast cancer risk in KPK. • Suggests limited utility of this SNP as a biomarker in this population.

10. Future Recommendations • Use larger sample sizes. • Study multiple SNPs in XRCC1 and related genes. • Consider environmental and lifestyle interactions. • Explore role in treatment response.

11. Acknowledgements • Mam Sana Khan (Supervisor), Dr. Hazrat Hussain (Co-Supervisor) • Faculty and IRNUM Hospital • My family and friends for their endless support • Grateful to Almighty Allah