Application of compounds that improve dicer expression in the preparation of medicines
A drug and low-expression technology, applied in drug combinations, medical preparations containing active ingredients, pharmaceutical formulas, etc., can solve problems such as unclear dynamic patterns of Dicer expression, and achieve the effect of improving the inhibitory effect and huge clinical promotion value
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[0137] a. Preparation of protein standard: Take a tube of BSA protein standard and dissolve it in 1.2ml of protein standard preparation solution to prepare a standard with a concentration of 25mg / ml, take out an appropriate amount of sample and prepare it to 0.5mg / ml as Use of standards for determination of protein concentration;
[0138]b. Add 0, 1, 4, 8, 12, 16, and 20 ul of standard to the 96-well plate, and add standard dilution to make the final volume 20 ul. The corresponding standard concentrations are: 0, 0.025 , 0.05, 0.1, 0.2, 0.3, 0.4, 0.5 mg / ml;
[0139] c. Add 20ul of protein sample to 96-well plate
[0140] d. Add 200ul of BCA working solution in the air where the samples and standards are added, and incubate at 37°C for 30 minutes. The formula of the working solution is: add BCA reagents A and B at a ratio of 50:1 to prepare an appropriate amount of BCA working solution;
[0141] e. Use a multifunctional microplate reader to measure the absorbance at A562.
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Embodiment 1
[0264] Example 1: Discussion on the relationship between Dicer enzyme and colitis and colon cancer
[0265] 1. The correlation and mechanism of Dicer expression with the pathogenesis of colitis and colon cancer
[0266] To investigate the expression of Dicer in precancerous colitis, paraffin-embedded colon samples from 56 patients with inflammatory bowel disease (27 patients with Crohn's disease and 29 patients with ulcerative colitis) and 57 normal subjects were examined. Dicer expression in paraffin-embedded tissue samples, immunohistochemical analysis showed that Dicer expression was reduced in inflamed colon tissue relative to normal tissue samples ( figure 1 A-B), in addition, the use of additional frozen inflammatory colon tissue from 46 inflammatory bowel disease patients and 34 normal population tissue samples for analysis showed that Dicer was reduced at protein levels, but not at mRNA levels ( figure 1 C-E); similarly, Dicer protein levels, but not mRNA levels, we...
Embodiment 2
[0279] Example 2: The effect of anastrozole on hydrogen peroxide-induced cell damage
[0280] Consistent with our previous study that Dicer overexpression promotes DNA repair, Dicer overexpression reduces H 2 O 2 Induced DNA damage, reduced H 2 O 2 Induces an increase in cytoplasmic DNA and alleviates H 2 O 2 Induced IL-6 expression ( Figure 12 A-C and Figure 13 A-B). By screening the drug library, we found that anastrozole can enhance the expression of Dicer ( Figure 14 A and Figure 15 A). Anastrozole-induced increase in Dicer expression, decreased response to H 2 O 2 susceptibility to induced DNA damage ( Figure 14 B-D and Figure 15 B-D). Therefore, treatment with anastrozole reduces H 2 O 2 Induced increases in cytoplasmic DNA and IL-6 expression ( Figure 14 E-F and Figure 16 A-B). Silencing Dicer expression partially abrogates the effects of anastrozole ( Figure 17 A-C and Figure 18 A-B). Together, these findings suggest that anastrozole may...
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