The Effect Of Recombinant Fn C-terminal Heparin-binding Domain Polypeptide On Metastasis Of Human Gastric Cancer SGC-7901 Cell Line

Objective:The aim of this Study was to evaluate the effect of Recombinant FN C-terminal heparin-binding domain polypeptide(rh FNHC36) on migration and metatasis of human gastric cancer SGC-7901 cell in vitro and vivo by exposing them to the different concentration of rh FNHC36,and to explore the probable mechanism.Methods:1.The crystal violet was used to evaluate the ability of adhesion of human gastric cancer SGC-7901 cell which was exposed to different concenrations of rh FNHC36(100μg/ml、200μg/ml、400μg/ml);2.The Transwell chamber was used to assessed the ability of migration and invasion of cells. 3.The expression of relevant integrin submits(αV、β1、β3)and EMT-related proteins(E-cadherin、Vimentin) was measured by Western Blot.4.The activity of matrix-metalloproteinase9(MMP9) in cell supernatant was analyzed by gelatin zymogra-phy.5.The human gastric cancer SGC-7901 cell transvascular metastasis model labeled with luciferase was use to evaluate the impact of rh FNHC36 on the cells’ ability to meta stasize in nude mouse.Results:1.In each group of rh FNHC36(100μg/ml、200μg/ml、400μg/ml),the cells’ adhesion rates to Matrigel Matrix were 81.04 ? 1.50%、74.77 ? 1.20% and 68.62 ? 2.37%,respectively,which were lower than the control group’s(it was assumed 100%).The differences were statistically significant(P<0.01).In each group of rh FNHC36(100μg/ml、200μg/ml、400μg/ml),the cells’ adhesion rates to FN were 80.63 ? 5.22%、79.00 ? 2.26% and 73.38 ? 2.02%,respectively.Compared with the control group(its adhesion rate was assumed 100%),only media and high dose group of rh FNHC36 had Statistical differences(P<0.05).2.In each group of rh FNHC36(100μg/ml、200μg/ml、400μg/ml),the numbers of migration SGC-7901 cells were 77 ? 15.29、65 ? 15.02 and 35 ? 6.42,respectively,which were less than the control group’s(130 ? 7.09,P<0.01).In each group of rh FNHC36(100μg/ml、200μg/ml、400μg/ml),the numbers of invasion SGC-7901 cells were 24 ? 3.78、17 ? 3.85 and 12 ? 4.74,respectively,and less than the control group’(48 ? 9.40,P<0.05).3.In each group of rh FNHC36(100μg/ml、200μg/ml、400μg/ml),the expression of integrin αV of SGC-7901 cells were 0.54 ? 0.14、0.45 ? 0.16 and 0.31 ? 0.08,respectively,and weaker than control group’s(0.94 ? 0.07,P<0.01).In e-ch group of rh FNHC36(100μg/ml、200μg/ml、400μg/ml),the expression of integrin β1 of SGC-7901 cells were 0.38 ? 0.13、0.35? 0.11 and 0.35 ? 0.12,respectively,wh-ich were weaker than control group’s(0.79 ? 0.03,P<0.01). However,rh FNHC36 has no effect on the expression of integrin β3、E-cadherin and Vime-ntine.4.In each group of rh FNHC36(100μg/ml、200μg/ml、400μg/ml),the activities of pro-MMP9 of SGC-7901 cells were 723 ? 49.42、622 ? 36.12 and 564 ? 38.19,respectively,and weaker than the control group’s(966 ? 49.78,P<0.01).In each group of rh FNHC36(100μg/ml、200μg/ml、400μg/ml),the activities of act MMP9 of SGC-7901 cells were 248 ? 23.16、191 ? 23.90 and 164 ? 22.28,respectivel y,which were weaker than the control group’s(308 ? 38.56,P<0.05).5.The total flux[unite:p/s] of tumor in liver of the rh FNHC36 group was 6.41E8 ? 6.45E8.Compared with the control group’s(1.48E10 ? 1.83E10),the differences were statistically significant(P<0.05).Conclusions: rh FNHC36 could inhibit gastric cancer cells SGC-7901’s ability of adhesion、migration and invasion in vitro and inhbit cells’ metastasis to liver in vivo,which may through down-regulating the expression of integrin αVβ1 and the activity of MMP9.