| In recent years,the number of the study that the relation ship between leukemia and angiogenesis is increasing. Large amounts of experiments suggest that the microvascular density ( MVD) in patients who suffered from leukemia is obviously higher than that in the normal people and many molecular factors that take part in angiogenesis. Most scholars think the VEGF is the most specific and having the most rapidly action factors among the molecular factors that participate in angiogenesis in tumor. The viro experiments have shown that the leuke-mic cell can secrete VEGF and express the functional receptor of VEGF, which suggest that the leukemic cell can maintain the growth and proliferation by au-tosecretion and in parasecretion indirectly which will stimulate the angiogenesis and secretion of other molecular from endothemia. Thalidomide once had been used to inhibit the pregnancy reaction, but it had been forbidden to use for it severe side - effect. In the near years, thalidomide have been found the new function of anti - tumor and be used to treat the multiple myeloma to receive the good prognosis. Thalidomide also be used singly or combined with chemotherapy to treat leukemia and it works. Thalidomide has the function of anti - tumor through inhibition of angiogenesis. Moreover, As2O3has been found to lower the expression of VEGF in leukemia cell and induce the apoptosis of cell.ObjectiveTo investigate the affect of thalidomide and As2O3 that of different dosage tothe secretion of VEGF and apoptosis of the cell of ANLL through the exviro culture of leukemic cell and study if there is coorperation in the thalidomide and As2O3.Method1. solation and culture of mononuclear cells in marrowHeparinized marrow of 2ml was taken under sterility, then it was putted into Ficoll - isopaque solution and was centrifuged (900g) to purified MNC, washed three times and cultured in the RPMI1640 medium containing 10% fetal bovine serum (FBS) , 100IU/ml penicillin and 100IU/ml streptomycin, placed in culture box with 5% CO2 saturation in 37C to proliferation.2. roupingChoosing the cells in logarithm growth phase, centrifuging, getting rid of upper liquid, adding cultural solution to abstaining the cellular density of 2 x10 /ml and then putting it into 24 - well plates.There are 6 groups that be held up. A: contrast group, B; 100ug/ml thalidomide, C: 300ug/ml thalidomide, D: 25 umol/1 As2O3 E: 100ug/ml thalidomide and 25 umol/1 As2O3 F: 300ug/ml thalidomide and 25 umol/1 As2O3 The samples of 6 groups were placed in cultural box with 5% CO2 saturation and in 37C for 72 hours.3. Checking apoptosis and the concentration of VEGFCollecting cellular suspension, centrifuging (1500r/min) for 5 min, staining with PI and checking the apoptosis cells with Fluid Cellular Instruments. Examing the concentration of VEGF in upper liquid with ELISA method. Taking the cells of before cultured and after cultured to smearing, Giemsa staining, observing the morphology of cells under the light microscope.Result1. The effect of drugs to the VEGF secretion of leukemic cells The effect of thalidomide to the VEGF secretion of leukemic cellsThe concentration of VEGF in upper liquid of A, B, C group are respectively 1640.19 287.10 pg/ml 1596. 88 +266.18 pg/ml 1242. 32 +281.95 pg/ml . There is no statistic difference between A group and B group. But there are obviously statistic difference between A group and C group, B group and c group ( P <0.05). The affect of As2O3 to the VEGF secretion of leukemic cellsThe concentration of VEGF in A group and D group are 1640.19 +287.10 pg/ml 933.99 +225.41 pg/ml. There is obviously statistic difference between the two groups (P <0.01).The affect of As2 O3 and thalidomide to the VEGF secretion of leukemic cellsThe concentration of E group and F group are 886. 36 250. 18 pg/ml and 605.86 208.51 pg/ml. There are obviously statistic difference with A, B and C group. And the concentration of VEGF is lower in F than in D group ( P = 0. 052).The affect of dr... |
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