Influence Of Silencing NF-kB P65 By RNA Interference On BSEP And NTCP Expression In Estrogen Effected Of Human Hepatic Cells

Object:To investigate the influence of silencing NF-κB by specific siRNA on expression of BSEP and NTCP in normal human hepatic cells by Hormone-Effected.Methods:1.Human hepatic cells are cultivated with estrogen effected, and then are transfected with NF-kB siRNA by using catiotic liposome lipofectamineTM 2000 as the transfecting agent. Cells transfected with negative control-siRNA, empty Liposome and nothing as control. Twenty-four hours later, cells are collected and prepared for Real-time PCR,Western blot analsis, and detrminnation of the migration30,6-Djhydroxycholanic inhibition rate.2. A comparative study of the protein expression on NF-κB in hepatic cells is done before and after estrogen effected twenty-four hours by Western blot.3. The mRNA and protein expression of BSEP and NTCP are detected in hepatic cells before and after siRNA tranfection by real-time PCR and Western blot at the same timeResults:1. BSEP and NTCP in normal human hepatic cells.2. The influence of estrogen effected expression of NF-κB in hepatic cells.The results are as following:The values in estrogen effected group is higher than that in blank control group(P<0.05). Date showed the obvious correlation between the protein expression of NF-κB and concentration of estrogen.3. The intervention of NF-κB p65 siRNA affects the expression of BSEP and NTCP in hepatic cells. Compared with normal control group, both mRNA and protein levels of NF-κB decrease obviously, and at the same time, cell migration inhibition rate increase obviously in NF-κB siRNA group. There is no significant difference between empty liposome group, negative control siRNA group and normal control group respectively(P>0.05).There is a significant positive correlation between the expression of BSEP,NTCP and siRNA P65(P<0.05). The value in NF-κB siRNA group is higher than that in empty liposome group and normal control siRNA group(P<0.05). But there is no significant difference between the empty liposome group and normal control siRNA group (P>0.05).Conclusion:1. Special of siRNA may regulate the expression of NF-κB in hepatic cells.2. The expression of BSEP and NTCP in hepatic cells can be up regulated via silencing of NF-κB by specific siRNA.3. Estrogen may regulate the expression of BSEP and NTCP in hepatic cells of human by NF-κB pathway and then affect the activity of hepatic cells of human.