Background: Oral squamous cell carcinoma is the most common oral cancer. It has a great predisposition to produce lymph node metastasis. Solid tumors are characterized by the presence of two major components: neoplastic cells and a specialized non malignant microenvironment in which they are immersed and are essential for survival and proliferation.. However the mechanism by which tumor microenvironment regulating this processes is unkown. Objective: The aim of the present work is to study the interaction between OSCC cells and their stromal microenvironment both in primary site and the secondary site in lymph node metastasis. This will be done through: Evaluation of the tumor microenvironmental major component: myofibroblast (CAF), tumor associated macrophage (M2, TAM) and lymphatic vessels, evaluation of Twist expression, an epithelial mesenchymal transition marker and correlate between the different studied parameters and clinicopathological grades of the tumor. Materials and Method:The present study carried out on 40 patients having OSCC. They classified into two groups according to lymph node metastasis. Antibodies used α-SMA, CD163, Twist and D2-40. Methods:  clinical evaluation from patients reports, paraffin sections from each case were employed for histological evaluation, haematoxylin and eosin for Broder and Anneroth evaluations, ASMA, CD163,Twist and D2-40. Results:The four markers showed significantly overexpression in OSCC with lymph node metastasis. α-SMA overexpression was related to advanced clinical stages, lymph node involvement and depth of tumor cell invasion. CD163 overexpression was related to tumor size, advanced clinical stages, lymph node metastasis and lymphoplasmatic infiltration. Twist overexpression was related to advanced clinical stage, depth of tumor invasion and lymph node metastasis. High mean of expression of D2-40 was found in advanced clinical stage, poorly differentiated OSCC and with increased depth of tumor invasion. The tumor cells invade the lymph node in the form of epithelial pearls , cell nests and small group of cells and with the same or higher histological grade of the primary tumor. All of the markers had reaction in lymph nodes. Conclusion: α-SMA,CD163,Twist and D2-40 overexpression might be an indicator of lymph node metastasis.