Simulated Ischemia And Reperfusion Injury Induce Hepg2 Cell The Expression Of Multidrug Resistance Associated Genes And Proteins

Objective: HCC is one of the most common malignant tumor in our coutry,its mortality only inferior to lung cancer.It is diagnosed already in middle or late period.chemotherapy is one of the important treatments for advanced HCC.Research shows that the majority of HCC are not sensitive to chemotherapy,Because of the high expression of multidrug resistance associated genes and proteins.Ischemia-reperfusion injury is clinically common damage factor.Removal of cancer tumor and transplantation is thus far the most efficient treatment for HCC,Hepatic blood flow occlusion is a common surgical skill,After block of liver blood supply,it can cause ischemia-reperfusion injury.If liver injury is serious,it will increase the risk of surgery and even cause liver failure.Studies have shown that liver cancer cells in hypoxia,low sugar environment,The expression of multidrug resistance associated genes and proteins can increase,which results in a decrease in chemotherapy effect,and even no effect,Hypoxia activates hypoxia-inducible factor 1.The study found that HIF-1 alpha has correlation with multidrug resistance associated genes, Silence or enhance the expression of HIF-1 alpha gene can reduce or enhance the expression of multidrug resistance associated genes. It suggested that HIF-1 alpha is one of the causes of multi-drug resistance of tumor cells. In order to further understand IRI and its effects on multidrug resistance,This experiment using Hepg2 cell model of IRI,This experiment using cell model of IRI,to explore IRI effect the expression of multidrug resistance associated genes and proteins,HIF-1 alpha of hepg2 cell.Discussing the association between HIF-1 alpha and multidrug resistance associated genes(MDR、LRP、MRP2). Methods:Establishing a Hepg2 cells model of ischemia-reperfusion injury.cell groups:blank control group and ischemia reperfusion intervention groups:ischemia 40 min,reperfusion respectively 0 h, 6 h, 12 h, 24 h, 48 h.Then observe the Hep G2 cell morphological changes after intervention,the RT-QPCR detects the expression of MDR1, MRP2, LRP and HIF-1 alpha,Western blot detects the expression of P- GP,MRP2, LRP,and HIF-1 alpha.The data statistics analysis by SPSS 17.0 software,The measurement data of multiple group were compared with single factor analysis of variance.α=00.5,The difference was statistically significant(P<0.05).Results: Cell morphological changes after IRI,dead cells increased obviously,Dilated intercellular spaces.The Intervention groups compared with blank control group:there are differences in the expression of MDR1 gene until IR-6h group(F = 116.16, P < 0.05),IR-48 h group was 3-fold grater than the blank control group. There are differences in the expression of MRP2 gene until IR-6h group(F= 133.81, P<0.05),IR-48 h group was 2.5-fold grater than the blank control group. There are differences in the expression of LRP gene until IR-6h group(F=146.64, P<0.05),IR-48hgroup was 3-fold grater than the blank control group.There are differences in the expression of HIF1- alpha gene until IR-6h group(F=113.01, P <0.05),IR-48 h group was 3-fold grater than the blank control group. there are differences in the expression of P-GP until IR-6h group(F=83.75, P < 0.05),IR-48 h group was 5-fold grater than the blank control group.There are differences in the expression of MRP2 protein until IR-6h group(F=52.70, P<0.05),IR-48 h group was 2-fold grater than the blank control group.There are differences in the expression of LRP protein until IR-6h group(F=2.01, P<0.05),IR-48 h group was 17-fold grater than the blank control group. There are differences in the expression of HIF1- alpha protein until IR-6h group(F=88.44, P < 0.05),IR-12 h group was 1.9-fold grater than the blank control group.Conclusion:1.The simulated ischemia reperfusion increased the expression of MDR1, MRP2, LRP of hepg2 cells and its corresponding protein P-GP, MRP2, LRP;2.The simulated ischemia-reperfusion induced the expression of HIF1 alpha gene and protein of hepg2 cells;3.We verified that HIF-1alpha can control MDR1 expression by the simulated ischemia-reperfusion...