Effect of naringin-mediated RhoA/ROCK1 pathway on oxaliplatin resistant human colorectal cancer cell line HT-29/L-OHP
Objective To investigate the effect of naringin-mediated Ras homologous gene family member A(RhoA)/Rho associated coiled coil binding protein kinase 1(ROCK1)pathway on enhancing the sensitivity of human colorectal cancer cell line HT-29 to oxaliplatin(L-OHP)chemotherapy.Methods Human colorectal cancer cell line HT-29 with a stable resistance to L-OHP(HT-29/L-OHP)was established.HT-29/L-OHP cells were inoculated to 96-well plates,with the cell density of 1.0×105/mL.Cells were induced with blank control(injection of sterile 0.9%NaCl 2 μL),5 μmol/L L-OHP 2 μL,100 μmol/L naringin 2 μL and 5 μmol/L L-OHP 2 μL+100 μmol/L naringin 2 μL,with 6 replicates.The cell viability and the half maximal inhibitory concentration(IC50)of each group were detected by methyl thiazolyl tetrazolium(MTT)assay.Real-time reverse transcription quantitative polymerase chain reaction(RT-qPCR)was used to detect the messenger RNA(mRNA)expressions of RhoA,Rock1,multidrug resistance associated protein 1(MRP1)and anti apoptotic factor(Bcl-2).The protein expressions of RhoA,Rock1,MRP1 and Bcl-2 were detected by Western blot(WB).Results Human colorectal cancer resistant cell line HT-29/L-OHP was successfully constructed.Compared with those treated with blank control or L-OHP group,cells treated with naringin or naringin+L-OHP had significantly lower cell proliferation ability,mRNA and protein expressions of RhoA,Rock1,MRP1 and Bcl-2,and IC50(P<0.05).There were no significant differences in the above indicators between the blank group versus L-OHP group,and between naringin group versus naringin+L-OHP group(P>0.05).Conclusion Naringin can significantly reduce the proliferation of L-OHP-resistant human colorectal cancer resistant cell line HT-29/L-OHP and improve its chemosensitivity to L-OHP by downregulating RhoA,Rock1,MRP1 and Bcl-2.
万方数据
维普
