Background and aim of the work: Previous studies verified that Endostatin, matrix metalloproteinase (MMP) -2 and -9, in addition to tissue inhibitors of metalloproteinase (TIMP) -1 may play a crucial role in prognosis of non-small cell lung cancer (NSCLC). In this study we will investigate the changes in the pretreatment serum levels of these factors and to evaluate their clinical implication in patients with advanced non-small cell lung cancer (NSCLC). Patients and methods: Pretreatment serum samples were collected from 25 patients and 10 control healthy individuals. The levels of Endostatin, MMP-2, MMP-9, and TIMP-1 were measured using a sandwich enzyme immunoassay kit. Results: The pretreatment serum levels of Endostatin and TIMP1 were significantly elevated and correlated with their stages and survival (P< 0.05), where, the serum level of Endostatin in healthy subjects was 81.20±23.99 ng/ml and in patients with NSCLC was 354.40 ± 164.01 ng/ml. The serum level of TIMP1 in healthy subjects was 1.49±0.29 ng/ml and in patients with NSCLC was 2.96±0.58 ng/ml. The serum level of MMP2 and 9 were non-significantly decreased in serum of NSCLC patients (P > 0.05), where the serum activity of MMP2 in healthy subjects was 0.14±0.03 ng/ml and in patients with NSCLC was 0.09±0.03 % and the serum activity of MMP9 in healthy subjects was 0.13±0.019 ng/ml and in patients with NSCLC was 0.10±0.03 %.  Conclusions: Our results indicated that the circulating levels of Endostatin, and TIMP-1 in patients with NSCLC may be valuable future tools for treatment planning and monitoring of treatment, however, these blood tests need to be standardized and validated in large-scale prospective clinical trials.