The Influence Of Compound26 To AO184 Cells And Its Signal Transduction Mechanism

Objective:1.To observe the influence to the migration and growth of AO 184 by Compound26.2.Discuss the function of RhoA passway to the migration and growth of AO184 induced by Compound26.Methods:1.Cell culture of AO184.2.Scratch assay.(1)The direct effect of Compound26.Randomly divided the AO 184 cells into 2 groups:the control group and the Compound26 group.Randomly divided the Compound26 group into 3 groups by the different concentration of Compound26:10uM?30uM?100uM,observe the migration status after 24h and 48h.(2)Activation assay:Randomly divided the cells into 3 groups:the control group,the Compound26 group and the activation group(with Ang? to activate RhoA),the activate group was Randomly divided into the Compound26 group,and the activate control group,to observe the migration status after 24h.(3)Inactivation assay:Randomly divided the cells into 3 groups:the control group,the Compound26 group and the inactive group(with exoenzyme C3 to inactive RhoA).Randomly divided the inactive group into the Compound26 group,and the inactivation control group,to observe the migration status after 24h.All the groups cultured in the same condition(37?,5%C02),to observe the migration status.3.Weste4nblot.Tested the change of the amount of RhoA protein in different groups.4.EMSA.Tested the change of theamount of transcription factor NF-kB in different groups.Results:(1)Compound26 could inhibit AO184 cells migration(30uM:16.8±3.6,100uM15.4±2.6)by the concentration of 30 uM after 24h.(P<0.05).Low concentration(10uM)of Compound26 could not inhibit AO 184 cells migration(18.6 ±2.5)(P>0.05).There was no difference between 30uM and 100uM of Compound26(P>0.05).Compound26 could inhibit AO 184 cells migration(30uM:45.0 ± 2.9,100uM:41.0 ± 2.8)by the concentration of 30 uM after 48h(P<0.05).Low concentration(10uM)of Compound26 could not inhibit AO 184 cells migration(50.2 ±3.6)(P>0.05).In the Compound26 100uM group,is was less compared to the Compound26 30uM group(P<0.05).(2)Cell migration:In the activation+Compound26 group,it was less(23.5±3.1)compared to the activation+control group(56.3±3.6)(P<0.05).(3)Cell migration:In the inactivation+Compound26 group,has no statistically significant difference(10.7±2.4)compared to the activation+control group(10.5±2.5)(P>0.05).(4)Westernblot:RhoA protein expression quantity:In the activation+Compound26 group,it was reduced compared to the activation+control group(P<0.05).In the inactivation+Compound26 group,it was not changed compared to the activation+control group(P>0.05).(5)EMSA assay:The quantity of activate NF-kB:In the activation+Compound26 group,it wasreduced compared to the activation+control group(P<0.05).Conclusion:Compound26 can inhibit the migration of AO184 cells.When RhoA is activated,Compound26 can play the inhibit role.When RhoA is inactived,Compound26 can not play the inhibit role...