A Distinct Role RhoB In Colorectal Cancer Suppression

Objective:To determine whether the liposome-mediated RhoB gene can inhibit the HCT116cells,and may therefore provide a potential therapy for human colon cancer.Methods:Cultured human colon cancer cell lines HCT116.There were five experiment groups:RhoB was up-regulated (pcDNA4-RhoB) and its negative control (pcDNA4-EGFP), RhoB was down-regulated (pGenesil-1-RhoB) and its negative control (pGenesil-1-HK), blank controller (HCT116). we transfected all of the plasmids into HCT116cells by liposome.In order to find out the expression of RhoB protein and the changes of HCT116cell lines,We analyzed the data by Western-Blot,MTT experiment and Flow cytometry.Results:(1)The vectors of pGenesil-l-RhoB,pGenesil-l-HK,pcDNA4-RhoB and pcDNA4-EGFP were transfected into human colon cancer cell lines HCT116by Lipofectamine2000respectively,fluorescence microscope was used to assure the transfection.(2)RhoB protein was detected in all of the five groups by western-blot. pcDNA4-RhoB-transfected samples exhibited a dramatic increase of RhoB protein compared to the pcDNA4-EGFP-only control and blank controller(P<0.05). pGenesil-1-RhoB-transfected samples exhibited no difference in RhoB protein compared to pGenesil-1-HK-only control and blank controller(P>0.05).(3)MTT experiment shows:Cell growth rate was slowed down obviously in pcDNA4-RhoB-transfected samples after the second day(P<0.05).(4)Using flow cytometry to examine cell’s apoptosis, The pcDNA4-RhoB-transfected samples exhibited a distinct difference in apoptosis rate compared to pGenesil-1-HK-only control and blank controller(P>0.05).Conclusion:RhoB may be useful for inhibiting growth and inducing apoptosis of HCT116.It may therefore provide a potential therapy for human colon cancer.Postgraduate student:Dong-dong Song (Surgery) Directed by Prof. Hai-bo Wang...