The Expression Of Calreticulin On Gallbladder Carcinoma Tissues And Its Effect On The Biological Behavior In Gallbladder Cancer Cells

Objective:To explore the expression of calreticulin(CRT)in gallbladder cancer tissue and its effect on the biological behavior in gallbladder cancer GBC-SD cells.Methods:Immunohistochemistry and RT-qPCR were applied to detect the protein and mRNA levels of CRT in gallbladder cancer,adjacent cancer and chronic cholecystitis tissues.Small interfering RNA was transfected into gallbladder cancer GBC-SD cells and Western blotting were used to detect the expression of CRT.The proliferation was determined by using cell counting kit-8(CCK-8)and clone assays.Flow cytometry were applied to detect the apoptosis and cell cycle.Migration was detected by wound healing and transwell assays,respectively.The expression of p-Akt and MMP-9 were detected by using Western blotting.Results:Expression of CRT in gallbladder cancer tissues is higher than adjacent cancer tissues and chronic cholecystitis tissues(t=5.571,P<0.05).The relative growth rate in the siCRT-1 ? siCRT-2 experimental group for 24 hours,48hourrs were(71.5±6.3)%,(79.5±2.7)%;(62.6±8.8)%,(55.6±2.6)%,respectively.The apoptosis rate in the blank group,the negative control group,siCRT-1 and siCRT-2 group were(3.0±1.8)%,(4.7±1.3)%,(13.6±1.0)%,(20.0±4.0)%,respectively.Would healing assays showed that the wound closure ratio in the blank group,negative control group,siCRT-1 and siCRT-2 group were(0.67±0.02),(0.58±0.02),(0.22±0.01),(0.37±0.04),respectively.Transwell experiments showed that the numbers of migration of GBC-SD cell in the blank group,negative control group,siCRT-1 and siCRT-2 group were(301.8±10.7),(296.6±14.5),(178.0±10.1),(165.2±12.0),respectively.compared with the blank group and the negative control group,the relative growth rate for 24 hours and 48 hours was significantly lower,the apoptosis rate was higher,the numbers of migration was lower((F=29.310,118.618,69.651,144.515,190.145,P<0.05).Compared with the blank group and the negative control group,the expression of p-Akt and MMP-9 were decreased after down-regulating the expression of CRT.Conclusions:The expression of CRT in gallbladder cancer tissue was higher.CRT downregulation mediated changes of biological behaviors in gallbladder cancer may be associated with p-Akt/MMP-9 signal pathway.