| Objective: The study was for purpose of evaluating the effect of Recombinant FN heparin-binding domain polypeptide(rh FNHN29 and rh FNHC36) on invasion and metastasis of human pancreatic cancer bxpc-3 cell, and exploring the probable involved mechanism.Methods and Contents: 1.The adhesion capacity of human pancreatic cancer bxpc-3 cell which was added to different concentrations of rh FNHC29 and rh FNHC36 was assessing by adhesion test; 2.The ability of mig-ration and invasion of bxpc-3 cell was assessing by Transwell chamber; 3.The expression of relevant integrin(?V, ?1) was analyzed by Western Blot; 4.ELISA was used to measure the concentration of matrix-metalloprotei-nase-9(MMP-9) in cell supernatant. 5.The human pancreatic cancer bxpc-3 cell transvascular metastasis model labeled with luciferase was use to evaluate the impact of rh FNHC29 and rh FNHC36 on the cells' ability to metastasize in nude mouse.Results: 1.In each treatment group of rh FNHN29(100?g/ml, 200?g/ml, 400?g/ml), the adhesion rates of bxpc-3 cell to FN were 89.44 ± 5.38%, 79.17± 3.56% and 75.96 ± 3.99% respectively, and in each treatment group of rh FNHC36(100?g/ml, 200?g/ml, 400?g/ml), the cells' adhesion rates to FN were 75.99 ± 4.34%, 69.69 ± 3.14% and 60.91 ± 0.31% respectively, were all lower than the cotrol group's adhesion rate(which was assumed to be 100%), the differences were statistically significant(P<0.05). In each group of rh FNHN29(100?g/ml, 200?g/ml, 400?g/ml), the cells' adhesion rates to Matrigel Matrix severally were 65.82 ± 3.75%, 58.44 ± 2.33% and 44.73 ± 3.39%, while in each group of rh FNHC36(100?g/ml, 200?g/ml, 400?g/ml), the cells' adhesion rates to Matrigel Matrix were 65.82 ± 3.75%, 58.44 ± 2.33% and 44.73 ± 3.39% severally, which were much lower than the control group's(its adhesion rate was assumed to be 100%) except the low dose group of rh FNHN29, the differences were statistically significant(P<0.05).2. In each treatment group of rh FNHN29(100?g/ml, 200?g/ml, 400?g/ml), the bxpc-3 cell numbers of migration were 21 ± 2, 13 ± 2 and 9 ± 2 respectively, and in each group of rh FNHC36(100?g/ml, 200?g/ml, 400?g/ml), the cell migration numbers were 19 ± 4, 11 ± 2 and 8 ± 2 respectively; compared with the control group(37 ± 8), the cells of treatment group that migrated through the polycarbonate membrane were obviously decreased, the deference had statistics significance(P<0.01). In each treatment group of rh FNHN29(100?g/ml, 200?g/ml, 400?g/ml), the bxpc-3 cell numbers of invasion severally were 23 ±3, 13 ± 3 and 9 ± 1, and in each group of rh FNHC36(100?g/ml, 200?g/ml, 400?g/ml), the cell invasion numbers severally were 23 ± 9, 14 ± 2 and 7 ± 3, were all less than the control group's(30 ± 7, P<0.01) except low dose group of rh-FNHN29 and rh FNHC36.3. In each treatment group of rh FNHN29(100?g/ml, 200?g/ml, 400?g/ml),the bxpc-3 cell's expression of integrin ?V respectively were 0.37 ± 0.02, 0.34 ±0.01 and 0.27 ± 0.01, and in each group of rh FNHC36(100?g/ml, 200?g/ml, 400?g/ml), the expression of integrin ?V respectively were 0.34 ± 0.02, 0.27 ± 0.01 and 0.17 ± 0.01, were all much weaker than control group's(0.47 ± 0.03, P<0.01). In each treatment group of rh FNHN29(100?g/ml, 200?g/ml, 400?g/m l), the bxpc-3 cell's expression of integrin ?1 were 0.38 ± 0.01, 0.33 ± 0.03 and 0.23 ± 0.02 severally, and in each group of rh FNHC36(100?g/ml, 200?g/ml, 400?g/ml), the expression of integrin ?1 were 0.43 ± 0.02?0.26 ± 0.01?0.19 ± 0.01 severally; compared with control group's(0.76 ± 0.04), the expression of integrin ?1 of treatment groups were much weaker, the deference had statistics significance(P<0.01).4. In each treatment group of rh FNHN29(100?g/ml, 200?g/ml, 400?g/ml),the concentration of matrix-metalloprotei-nase-9(MMP-9) in bxpc-3 cell supern-atant respectively were 94.17 ± 0.92pg/ml, 85.24 ± 2.97pg/ml and 71.10 ± 6.95pg/ml, and in each group of rh FNHC36(100?g/ml, 200?g/ml, 400?g/ml), the concentration of matrix-metalloprotei-nase-9(MMP-9) in cell supernatant were 77.86± 7.16pg/ml, 49.95 ± 1.59pg/ml and 35.91 ± 2.80pg/ml respectively, which were all much lower compared with control group's(121.99 ± 2.79pg/ml), the differences were statistically significant(P<0.01).5. The total flux [unite:p/s] of tumor in liver of the rh FNHN29 and rh FNHC36 group respectively were 5.97E7?3.52E7 and 6.99E7?4.70E7, which were much lower compared with the control group's(4.96E8?2.52E8), the differences were statistically significant(P<0.01).Conclusions: Either of rh FNHN29 or rh FNHC36 could inhibit pancre atic cancer bxpc-3 cell's ability of adhesion?migration and invasion in vitro an d inhbit cells' metastasis to lungs in vivo probably by down-regulating the exp ression of integrin ?V?1 and the activity of MMP-9 pr-obably. |
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