| Objective:Hepatic cell carcinoma (HCC) is one of the world’s six major malignancies, ranked third in the common male malignancy in women ranked eighth.HCC with multi-vessel, high degree of malignancy, fast growth,a wide range of transfer, high recurrence rate and other characteristics, clinical treatment effect is not ideal, often in need of postoperative adjuvant chemotherapy, but due to liver cancer chemotherapy often multidrug resistance (multidrug resistance, MDR) phenomenon, a serious impediment to the effects of chemotherapy.SUMO protein know to be associated with MDR.SUMOylation a important post-translational modification form of c-Jun, P53and HIF-1and other protein is,which affect the physiological and biochemical function.SUMOylation of some proteins may changed when the process of liver cancer formed resisitant,that may impact proteins Physiological function, so that the chemotherapy drug resistance increased.In this study, preliminary study whether SUMOylation play a role in liver cancer drug resistance and how this phenomenon occurrence.Method:1. SUMO protein expression in Cancer tissues and adjacent tissues detected by immunohistochemical method.2.We use5-fluorouracil (5-FU) induce5-FU resistant cell line. By compare morphology, growth curve, and sensitive to other chemotherapy drugs of Hep G2and HepG2/R cells, to prove whether we get the multidrug resistance (MDR) cell lines HepG2/R.3.Compare SUMO-1protein and various enzymes in SUMOylation between Hep G2and Hep G2/R by western blot and to determine wherther liver cancer drug resistance associated with SUMOylation and how this phenomenon occurs.Result:1.Immunohistochemical results showed that SUMO-1of the20cases of cancer tissue samples are highly expressed than adjacent tissues,exit in cytoplasm and nucleus.2.We successfully established multi-drug-resistant hepatoma cell line (HepG2/R) by take advantage of5-FU through one year.There is differece between HepG2/R and HepG2in morphology,in Hep G2/R pseudopodia extending.There is no significant difference between the two types of cells in the cell proliferation ability.When the5-FU concentration in0.8-6.4μg/ml, and its inhibitory effect on HepG2/R significantly lower than HepG2.In addition, HepG2/R cross-resistance to paclitaxel and oxaliplatin.3.When different concentrations of5-FU paly role in the two cells,SUMO protein in HepG2and HepG2/R cells were significantly different.HepG2/R cells express specific binding of SUMO protein and free SUMO protein in60kDa and11kDa.Meanwhile, in the the30kDa position SUMO specific binding protein expression was significantly increased.4.Compare changes in enzymes involved in the SUMO modification process.The two cells AOS-1expression was no significant difference, Uba-2expression in the dose of5-FU concentration in HepG2/R cells.Senp-1expression in HepG2/R cells were significantly increased, and in dose of5-FU concentration.Conclution:1.We successfully established multi-drug-resistant hepatoma cell line (HepG2/R) by take advantage of5-FU through one year.Hep G2/R apparent resistance of5-FU by drug sensitivity test.In addition, HepG2/R cross-resistance to paclitaxel and oxaliplatin.The morphology of resistant hepatic cancer cell is changed.By tracing the growth curve of Hep G2cells and Hep G2/R cells found that there was no difference in the prolifetation.2.Our study found that liver cancer multidrug resistance significantly associated with SUMOylation,this phenomenon was mainly caused by changes of Uba-2, and Senp-1expression.Therefore, to prevent changes in the SUMOylation state could become an effective measure for the treatment of liver cancer... |
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