| Objective(1)This experiment is to study expression of vWF and integrinβ3 in human lung cancer cell line H460 and the association between the two factors and human lung cancer metastasis.The H460 manifested higher metastasis potential than the CLC-82 line.(2)To observe vWF adhesion with the human lung cancer cell line H460 and GLC-82.(3)To observe the correlation of vWF and integrinβ3.Methods(1)vWF and integrinβ3 protein was examined using immunohistochemical staining method in H460 and 6LC-82 human lung cancer cells.(2)Adhesion capability of human lung cancer cells H460 and GLC-82 on vWF was examined using vWF adhesion experiment and MTT.(3)Correlation of vWF and integrinβ3 in tumor metastasis was examined using vWF adhesion experiment,antibody inhibition experiment and MTT.Results(1)Expression of vWFand integrinβ3 in human lung cancer cells H460 was detected strongly positive immunoreactivity,in GLC-82 was both weak expression.(2)Expression of integrinβ3 of human lung cancer cells H460 and 6LC-82 was not changed significantly blocking by anti-vWF. Expression of vWF of human lung cancer cells H460 and GLC-82 was not changed significantly blocking by anti-integrinβ3.(3)Human lung cancer cells H460 and GLC-82 were able to adhere to vWF in dose-dependent.Adhesion of H460 and GEC-82 on vWF increased,when increasing vWF concentration from 0.5μg·ml-1,1.0μg·ml-1to 2.0μg·ml-1.Adhesion did not increased after vWF 2.0μg·ml-1.(4)When increasing anti-integrinβ3 concentration from 70μg·ml-1,75μg·ml-1,80μg·ml-1to 85μg·ml-1we observed adhesion of H460 cells on vWF was inhibited blocking by anti-integrinβ3,which A Value decreased from 1.5923±0.0561 to 0.5507±0.0309,P=0.01619.When increasing anti-vWF concentration from 1.0μg·ml-1,1.5μg·ml-1to 2.0μg·ml-1,we observed adhesion of H460 cells on vWF was inhibited too blocking by anti-vWF,which A Value decreased from 1.6023±0.0601 to 0.5380±0.0340,P=0.01598.Blocking adhesion of H460 cells on vWF by anti-integrinβ3 had the same effect with blocking by anti-vWF.We found no significant differences between the two antibodyes inhibiting adhesion of H460 cells on vWF.(5)When increasing anti-integrinβ3 concentration from 70μg·ml-1,75μg·ml-1to 80μg·ml-1,we observed adhesion of GLC-82 cells on vWF was inhibited blocking by anti-integrinβ3,which A Value decreased from 1.3969±0.0480 to 0.3887±0.0237, P=0.01519.When increasing anti-vWF concentration from 1.0μg·ml-1to 1.5μg·ml-1,we observed adhesion of GLC-82 cells on vWF was inhibited too blocking by anti-vWF,which A Value decreased from 1.3968±0.0490 to 0.3510±0.0271,P=0.01609.We found no significant differences between the two antibodyes inhibiting adhesion of GLC-82 cells on vWF.Conclusions(1)Human lung cancer cells H460 and GLC-82 both express vWF and integrinβ3,and the expression in H460 cells,which manifest higher metastasis potential,is higher than in GLC-82 cells.So vWF and integrinβ3 increast human lung cancer metastasis.(2)Human lung cancer cells H460 and GLC-82 are able to adhere to vWF.Adhesion to H460 cells are higher than to GLC-82 cells.So vWF contributes to tumor metastasis by adhering to cancer cells.(3)Anti-integrinβ3 and anti-vWF can inhibite vWF adhesion to human lung cancer cells H460 and GLC-82.So,maybe integrinβ3 is the receptor of vWF. |
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