The Effects Of Bortezomib And Its Combination With5-FU On The Proliferation And Apoptosis Of The Choriocarcinoma Cells

Choriocarcinoma is one of the most serious forms of gestational trophoblastic tumor, which occurs in women of period of duration. Clinical metastasis is very common and it is characterized by early hematogenous spread. Chemotherapy is the preferred treatment to choriocarcinoma. it can be cured by using combination chemotherapy.5-fluorouracil and methotrexate are commonly used in clinical treatment of choriocarcinoma, however, drug resistance and relapse are found in some patients. By increasing the dose of conventional chemotherapy drugs, we can get some efficacy, but the side-effects cause severe harm to the patient’s physical and psychological healthy. So it is very necessary and important to look for new chemotherapeutic agents for optimizing treatment protocols, minimizing side-effects and inereasing the sensitivity to chemotherapy.Bortezomib, a new UPP(ubiquitin-proteasome pathway) inhibitor, is a novel anti-tumor targeted therapies drugs, it has been used in the treatment of recurrence and refractory multipl myeloma in clinic. Bortezomib had shown a promising antitumor activity in a number of preclinical cancer cell-line models in vivo and vitro, but its anti-tumor effects of choriocarcinoma cell is unclear. The purpose of this study is to investigate the effect of proteasome inhibitor bortezomib alone and combination with5-FU on proliferation, apoptosis of JEG-3cells and the expression of NF-κB.In this research, the human choriocarcinoma cell line, JEG-3, were randomly divided into bortezomib group(0.1-200nmol/1),5-FU group(0.025-200μg/ml), bortezomib+5-FU group(bortezomib10nmol/1 combinding with5-FU0.025μg/ml、0.25μg/ml、2.5μg/ml、25μg/ml), control group(Onmol/1group). CCK-8assay was used to test the inhibition rate of cell proliferation. Annexin V/propidiumiodide(PI) staining and Flowcytometer (FCM) was adopted to analyze the apoptosis rate. Optical microscopy and fluorescence microscopy was used to observe changes in cell morphology.immunofluorescence method was used to detect the expression of NF-κB. The results showed that compared with the control group, the inhibition rates of cell proliferation in bortezomib group(4.43±0.02)%～(71.31±0.54)%were significantly higher(P<0.05), and the effect was correlated with action time and drug concentration. The apoptosis rate of the bortezomib group (16.61±0.81)%was also significantly higher than that of the control group (P<0.05). JEG-3cells was treated with10nmol/1bortezomib for48hours, the expression of NF-κB protein was increased by immunofluorescence. Compared with the control group, the inhibition rates of cell proliferation in5-FU group (12.70±01)%～(75.35±0.57)%were significantly higher(P<0.05). The inhibition rates(60.16±0.40)%(67.00±0.29)%and apoptosis rates (19.44±0.38)%of cells in bortezomib+5-FU group were more significantly higher than those in5-FU group or bortezomib group(P<0.05).CDI of cytotoxicity of bortezomib in combination with5-FU was all lower than0.85.As shows above, it can safely draw the conclusion that Bortezomib can inhibit the proliferation of JEG-3cells, and induce apoptosis by increasing the expression of NF-κB. Bortezomib combination with5-FU can inhibit the proliferation of JEG-3cells and induce apoptosis, they had synergism in cytotoxicity to JEG-3cells.