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The Influence Of TNP-470 And/or IL-2 On The Adhesion Of Lung Cancer Cells To Endothelial Cells And The Expression Of Adhesion Molecule And MMP

Posted on:2003-05-09Degree:MasterType:Thesis
Country:ChinaCandidate:B ZhouFull Text:PDF
GTID:2144360092496105Subject:Pathology and pathophysiology
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InstructionMalignant tumor cells have the capability of local invasiveness and distant metastasis and the ability of inducing angiogenesis is the prequisite of this effect. Commonly speaking, tumor metastasis includes five steps; neoplasm angiogenesis, neoplasm cells adhere to ECs, local basement membrane is degraded and destructed, neoplasm cells migrate to the secondary sites and the neoplasm cells proliferate on the secondary sites. So we can conclude that neoplasm angiogenesis plays a key role in the process of metastasis. Therefore discussing angiogenesis inhibitors effect on neoplasm metastasis may direct clinical treatment.TNP - 470 is a specific angiogenesis inhibitor, and it can inhibit the proliferation of ECs. The study in vivo indicates that it can inhibit many neoplasm tissues growth and metastasis. At present, the explanation of TNP -470S effect mainly focuses on the inhibition of neoplasm metastasis. The mechanism has been studied from multiple levels. In this experiment , we discuss the effect of TNP - 470 on the adhesion of lung cancer cells to ECs and the expression of VCAM - Ion ECs, CD44V6, MMP - 2, MMP - 3 on lung cancer cells in order to elucidate the mechanism that TNP - 470 inhibits neoplasm metastasis. Mo-reover, whether the combinative effect of TNP - 470 and IL - 2 is stronger than single treatment.MethodsFirstly, HUVECs were cultured in 96 - well plates by modified Jaffe method. The neoplasm cells were added to the culture when ECs were confluent, at the same time,divide the ECs into 12 groups;negative control, IL - 2 (1000U/ml) , 10 ~6 g/mlTNP - 470; 10-6 g/mlTNP -470 + IL - 2; 10"8g/mlTNP -470; 10 "8g/mlTNP - 470 + IL - 2; 10 -10g/ml TNP -470; 10 -10g/nuTNP -470 + IL -2; 10 -12g/rnlTNP _ 470; 10 ~12 g/mlTNP - 470 + IL - 2; normal control; positive control. Test the adhesion rate of neoplasm cells to ECs by MTT.Culture ECs and neoplasm cells on the glass slides and examine the expression of VCAM - 1 on ECs and CD44V6 ,MMP -2,MMP -3 in the neoplasm cells by immunhistochemistry and image - analysis. Then analyze the results statistically.ResultsThe adhesion rate of neoplasm cells to ECs is 72. 7+5.4% whenthere is no treatment. In the concentration range of 10-12g/ml-10-6 g/ml,TNP -470 can decrease the adhesion rate of these two cells dose - dependently ( P < 0. 05 ). When treated with IL - 2 ( 1000U/ ml) , the adhesion rate is decreased significantly compared with the control( P < 0. 05 ) . Combinative treatment of various concentrations of TNP - 470 and IL - 2 is stronger than single treatment respectively ( P <0. 05). The results of immunhistochemistry indicate that with the concentration of TNP - 470 increasing , the positive rate of VCAM - 1 on ECs and CD44V6 on lung cancer cells is decreased( P < 0.05 ). Butthe result of the expression of MMP - 2, MMP - 3 in the neoplasm cell plasma is not changed significantly with the changing concentration of TNP - 470 by image - analysis( P > 0. 05 ).DiscussionNeoplasms growth and metastasis is vascular - dependent. TNP -470 is a specific angiogenesis inhibitor and it can inhibit many kinds of neoplasm growth and metastasis. Its main target is normal EC cycle. TNP -470 can block normal EC into G1 phase , so the percentage of G0 phase cells is increased. Moreover, it has also been reported TNP - 470 can stimulate host immune system, which can make neoplasm cells decrease or apoptosis. IL - 2 is a widely ?used cytokine that treats neoplasm. It is the most important and potent T cell growth factor and it suppresses tumor through complicated immune regulation. However, high concentration of IL - 2 has cytotoxic effect which limits its clinical use. There are some reports of combinative treatment of TNP - 470 and other models, but the effect of combinative treatment of TNP -470 and IL -2 has not been reported. So-the aim of this experiment is to discuss whether TNP - 470 and/or IL - 2 inhibits the adhesion of neoplasm cells and ECs, and the combinative effect of these two drugs is stronger than single treatm...
Keywords/Search Tags:TNP-470, IL-2, EC, adhesion, neoplasm cell
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