Effect Of SCRIB On The Malignant Biological Behavioe Of Human Hepatocarcinnoma Cell Line Smmc-7721 Cells

Objective:(1)To investigate the effects of SCRIB on the malignant biological behaviors such as proliferation,apoptosis,invasion and migration of human hepatocarcinoma cell line smmc-7721;(2)To analyze the correlation between SCRIB and RAS apoptosis signal c-Jun N-terminalkinase(JNK)and explore preliminary mechanism of promoting apoptosis of SCRIB.Methods:(1)The experiment was divided into three groups: Untreated human hepatocarcinoma cell line smmc-7721 group(Control group),empty plasmid human hepatocarcinoma cell line smmc-7721 group(negative control group,NC group),recombinant plasmid SCRIBpcDNA3.1-3xFlag-C human hepatocarcinoma cell line smmc-7721 group(Scrib group).(2)SCRIBpcDNA3.1-3xFlag-C was transfected into human hepatocarcinoma cell line smmc-7721 by liposome-mediated method.(3)Western blot was used to detect the expression of SCRIB gene and to determine whether SCRIB was successfully transfected.(4)The effects of SCRIB on proliferation,apoptosis,migration and invasion of human hepatoma smmc-7721 cells were detected by flow cytometry,cell scratching and cell invasion assays.(5)The expression of JNK protein in three groups of cells was detected by Western Blot.Results:(1)The relative expression of Scrib protein in Scrib group was significantly higher compared to that of Control group and NC group(P<0.05).There was no significant difference of in relative expression of Scrib protein in Control group and NC group(P>0.05).(2)The results of flow cytometry detection of 24 H transient cell cycle showed that the percentage of G1 phase smmc-7721 cells was lower and the percentage of S phase smmc-7721 cells was higher in the Scrib group than in the Control group and the NC group(All P<0.05).There was no significant difference in the percentage of cells between G1 phase and S phase in the NC group(P>0.05).(3)The apoptosis rate was significantly higher in Scrib group than Control group and NC group(P<0.05);There was no significant difference in apoptosis rate between Control group and the NC groups(P>0.05).(4)The results of cell scratching test showed that the scratch repair rate of Scrib group was lower than Control group and NC group(P<0.05).There was no significant difference in scratch repair rate between Control group and NC group(P>0.05).(5)The results of cell invasion assay showed that the invasive ability of Scrib group was lower than Control group and NC group(P<0.05).There was no significant difference of cell invasion ability between Control group and NC group(P>0.05).(6)The relative expression of JNK protein in Scrib group was significantly higher than in Control group and NC group(P<0.05).There was no significant difference in JNK protein expression between Control group and NC group(P >0.05).Conclusion:(1)SCRIB can promote apoptosis,inhibit proliferation,migration and invasion in human hepatocellular carcinoma smmc-7721 cells and may become a new target for liver cancer treatment.(2)SCRIB promotes apoptosis of hepatoma cells,which may be related to the expression of JNK protein and the promotion of RAS apoptosis signaling pathway.