| Osteosarcoma is malignant bone tumor and because of the introduction of neo-adjuvant chemotherapy, the 5-year survival rates of the patients have been improved from 10%-20% to 60-70% in the past two decades. At present, Cisplatin, Adriamycin Methotrexate Cyclophosphamide are the routine chemotherapeutics. Though the overall survival rates have been improved obviously, some patients are resistant to chemotherapy and their survival rates are still low. Meanwhile some adverse effects are often seen when high dose chemotherapy performed. So the doses and the treatment effects are limited by the adverse effect in someway when using one drug for chemotherapy. Searching for new drugs or chemosensitizers for osteosarcoma chemotherapy becomes more popular these years. Suramin is a synthesized drug for the treatment of Africa trypanosomiasis. It has appeared some growth inhibition or apoptotic effects on lung cancer prostate cancer, mammary cancer and so on, which may be related to the disruption of the binding of growth factors to its receptors non-specifically and the inhibition the activity of some enzymes. Researches have demonstrated that suramin could suppress the activities of alkaline phosphatase (AKP) and telomerase in osteosarcomas in osteosarcoma. And suramin was also found to have the ability of inhibiting the growth of osteosarcoma stimulated by growth factors. Our research is aimed to investigate the cytotoxity of suramin on Osteosarcoma MG-63 alone or combined with cisplatin, which will provide some experimental proofs of suramin in the treatment of osteosarcoma.Materials and Methods一, Materials:Human Osteosarcoma MG-63 cell line (CCTCC); Cisplatin (China); Suramin (Sigma); MTT (Sigma); MEM culture medium (Gibcol); DMSO (Sigma); Fetal Calf Serum (Hangzhou); 96-well plate (Biochemicals); Cell culture incubator (Heraeus); Inverted microscope (Axio-avert); Elisa Reader (Opsys MR DYNEX). 二, Methods:1 Human Osteosarcoma cell MG-63 were cultured in MEM culture medium containing 10% (vol/vol) FCS(which has been inactivated in 56℃ for 30 min) 100 U/ml penicillin and 100 U/ml streptomycin at 37C in a CO2 incubator, 95% air 5% CO2 and at 100% humidity. A solution containing 0.25% Trypsin and 0.02% EDTA was used for detaching MG-63 after 48h for subculture.2.1 MG-63 cells were diluted with MEM culture medium to 1X105 cells/ml and each well of the 96-well plates was filled with 100ul solution which containing 104cells. After incubation for 24h, the cells attached to the bottom of the well and covered about 60% of the area. Then each well received another 100ul MEM solution which containing drugs or medium only. The five wells of the same group would have an identical concentration of cisplatin or suramin. The final concentrations of the drugs were ranged from 0.625ug/ml to 5 u g/ml for cisplatin and from 50 u g/ml to 400 u g/ml for suramin.2.2 The middle two concentrations of the two drugs were combined crossly to form four groups which containing the two drugs. Then all the cells in the plates were incubated for another 48h.The absorbance (A) value of each well was read at the wave-length of 570nm on an Opsys MR Elisa reader. Cytotoxity index (CI) was calculated, CI=(control group A-testing group A)/control group A X 100%.2.3 MG-63 cells were cultured in glass with culture medium only for about 24 hand changed the medium with suramin at concentration of 100 u g/ml, 200 u g/ml,cultured for further 72h and the morphologic changes were observed by invertedmicroscope.三, Statistical method: The numerical results were analyzed with SPSS-11.0 statistics program.RESULTS1 . The mean absorbance (A) value (read at 570nm) and CI value of MG-63 cells incubated with cisplatin for 48h, see table 1:* the value has statistical difference compared with control group and p<0.05 the value has statistical difference compared with control group and p<0.012 The mean absorbance (A) value (read at 570nm) and CI value of MG-63 cells incubated with suramin for 48h, see table...
|
PDF Full Text Request