Application of phosphodiesterase4 inhibitor ZL-n-91 in preparing medicine for treating prostate cancer proliferation and transfer
A phosphodiesterase, prostate cancer technology, applied in the field of tumor biology, to achieve the effect of inhibiting proliferation and migration, high selectivity, and good development and application prospects
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Embodiment 1
[0026] Example 1: The effect of ZL-n-91 on the proliferation of prostate cancer cells was detected by CCK8 method.
[0027] 1) Take PC-3 cells in logarithmic growth phase and prepare single cell suspension. 100ul cell suspension per well (containing 1×10 4 cells) were inoculated in a 96-well plate, and were divided into 5 groups, blank control group, 10uM, 50uM, 100uM, and 200uM groups, with 6 duplicate holes in each group, and the cells were pre-cultured for 24 hours (at 37 ° C, 5% CO 2 Under conditions);
[0028] 2) Replace the fresh medium, add different concentrations of ZL-n-91 to each group, and continue to culture the cells for 24 hours and 48 hours respectively (at 37 ° C, 5% CO 2 Under conditions);
[0029] 3) Add 100ul of 10% CCK-8 solution to each well to avoid bubbles;
[0030] 4) Continue to incubate the cells for 1-2 hours, take out the culture plate at 30 min, 60 min, and 90 min respectively, and measure the absorbance at 450 nm with a microplate reader.
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Embodiment 2
[0032] Example 2: Treatment of nude mice with subcutaneous PC-3 tumors.
[0033] 1) Take the PC-3 cells in the logarithmic growth phase, prepare a single cell suspension with serum-free F-12K medium, and use 120ul cell suspension per tube (containing 2×10 6 cells) in 1.5ml EP tubes;
[0034] 2) Draw 0.1ml of the prepared cell suspension with a 1ml sterile syringe and inoculate it subcutaneously in nude mice;
[0035] 3) The mice were given drug treatment 3 days after inoculation.
[0036] 4) Divide nude mice into two groups: solvent control group and administration group (10mg / kg), intragastric administration of drug treatment every day, and measure subcutaneous tumor volume and mouse body weight of nude mice twice a week;
[0037] 5) When the mouse tumor volume reaches 1500mm 3 , the mouse tumors were taken out, the tumor sizes were compared, and the tumor weights were recorded.
[0038] The result is as figure 2 showed that the administration of drug treatment had no eff...
experiment example 3
[0039] Experimental Example 3: ZL-n-91 inhibits the expression of Ki-67 in transplanted tumor tissues.
[0040] The tumor tissues mentioned above were taken, fixed overnight with 4% formalin, embedded in paraffin and sectioned for tumor proliferation antigen Ki67 staining.
[0041] The result is as Figure 5 As shown, the positive rate of Ki67 staining of the tumor tissue in the administration group was significantly lower than that in the blank group, indicating that the proliferation of tumor cells in the administration group was reduced.
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