ABSTRACT
BACKGROUND: One of the most characteristic features of malignant tissue is the high level of intratumoral hypoxia, which is considered as a powerful factor for induction of tumor aggressiveness and malignant progression. Pancreatic cancer (PC) is a near fatal disease with very unfavorable clinical outcome despite the application of different treatment regimes. It was shown that PC is characterized by high level of hypoxia. AIM: To clarify the correlation between tumor hypoxia and ADAM8 protein expression. MATERIALS AND METHODS: ASPC-1, Panc-1, BxPC-3, Capan-1, MiaPaCa-2, Colo-357, Su8686 and T3M4 cell lines were used in the study. Expression of mRNA ADAM8 was evaluated by real-time quantitative polymerase chain reaction method. Immunoblot analysis was used to evaluate the expression of ADAM8 protein. RESULTS: Hypoxia induced a 2.5-5.9-fold increase of ADAM8 mRNA levels of in the examined pancreatic cancer cell lines except Panc-1 (p=0.046). On the protein level, hypoxia induced a 1.2-5.9-fold increase of the ADAM8 prodomain removal form (90 kDa) in 5/8 pancreatic cancer cells. Moreover, hypoxia induced a 1.3-2.0-fold increase of the remnant form ADAM8 (60 kDa) in 4/8 pancreatic cancer cell lines: Aspc-1, Colo-357, Panc-1, T3M4. CONCLUSION: It was observed the clear tendency in the increase both of ADAM8 mRNA and ADAM8 protein levels in pancreatic cancer cell lines under hypoxia compared to normal conditions of oxygenation. A potential role of ADAM8 as a hypoxia-dependent protein in the pathogenesis and evolution of pancreatic cancer that is characterized by high level of intratumoral hypoxia can be suggested.
Subject(s)
ADAM Proteins/biosynthesis , Gene Expression Regulation, Neoplastic , Hypoxia , Membrane Proteins/biosynthesis , Pancreatic Neoplasms/metabolism , Cell Hypoxia , Cell Line, Tumor , Humans , Immunoblotting , Neoplasm Invasiveness , Oxygen/chemistry , RNA, Messenger/metabolism , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
AIM: To investigate the relationship between tumor hypoxia in vivo, activity of matrix metalloproteinases (MMPs), and metastatic potential of tumor. MATERIALS AND METHODS: Lewis lung carcinoma (3LL) was used in this study. Total activity of MMP-2 and -9 in tumor was measured biochemically, tumor hypoxia level was assessed by (31)P NMR spectroscopy in tissue perchloric extracts. RESULTS: It was determined that hypoxia level in primary tumor has been concomitantly increasing along with tumor growth and correlated with metastasis level in lung. The positive correlation between hypoxia level and activities of MMP-2 and MMP-9 in primary tumor was registered. Moreover, the activity of MMP-2 and -9 in 3LL (primary tumor) directly correlates with metastasis level in lung. CONCLUSION: This study demonstrated that the growth of primary tumor is distinctly accompanied by an increase of tumor hypoxia level which positively correlates both with the activity of MMP-2 and -9 in primary tumor and metastatic efficiency.
