Background: Breast lesions are a significant global health concern, often requiring invasive procedures for definite diagnosis. Cytological and molecular markers present alternative, potentially less intrusive diagnostic methodologies. This study aims to compare the effectiveness of cytological and molecular markers for the diagnosis of breast lesions. Methods: A cross-sectional study was performed on a cohort of 500 patients presenting with breast lesions, undergoing cytological evaluation (Fine-Needle Aspiration Cytology) and molecular marker testing (HER2/neu, ER, PR, and Ki-67). Concordance between the two methods was evaluated. Results: Both cytological and molecular marker evaluations showed significant diagnostic potential. There was substantial concordance between the two modalities. However, molecular markers exhibited a slightly higher diagnostic sensitivity and specificity, especially in the detection of pre-malignant and malignant lesions. Conclusion: Cytological and molecular markers each offer valuable, complementary diagnostic approaches. While cytology provides rapid, cost-effective results, molecular markers offer increased sensitivity and specificity. Their combined usage can help minimize invasive procedures and improve patient outcomes. Further research is recommended to refine the usage of these diagnostic tools.