Application of compounds with enhanced Dicer expression in drug preparation
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
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preparation example Construction
[0137] a. Preparation of protein standard: Take a tube of BSA protein standard and dissolve it in 1.2ml protein standard preparation solution to prepare a standard with a concentration of 25mg / ml. Use of standards for determining protein concentration;
[0138]b. Add 0, 1, 4, 8, 12, 16, and 20ul of standard substances into the 96-well plate, and add the standard substance diluent to make the final volume 20ul, and the corresponding standard substance concentrations are: 0, 0.025 , 0.05, 0.1, 0.2, 0.3, 0.4, 0.5mg / ml;
[0139] c. Add 20ul protein sample to 96-well plate
[0140] d. Add 200ul of BCA working solution to 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 reagent A and B at a ratio of 50:1 to make an appropriate amount of BCA working solution;
[0141] e. Use a multifunctional microplate reader to measure the absorbance at A562.
[0142] f. Draw a protein standard curve according...
Embodiment 1
[0264] Embodiment 1: Dicer enzyme and colitis and the exploration of colon cancer relation
[0265] 1. The correlation and mechanism between the expression of Dicer and the pathogenesis of colitis and colon cancer
[0266] In order to study 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 tested. Dicer expression in paraffin-embedded tissue samples, immunohistochemical analysis showed that in inflammatory colon tissues, compared with normal tissue samples, Dicer expression decreased ( figure 1 A-B), in addition, using the frozen inflammatory colon tissues of 46 inflammatory bowel disease patients and 34 normal population tissue samples for analysis, the results showed that Dicer was reduced at the protein level, but not at the mRNA level ( figure 1 C-E); similarly, Dicer protein levels, but no...
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 reduced H 2 o 2 Induced DNA damage, reduced H 2 o 2 induced an increase in cytoplasmic DNA and alleviated the H 2 o 2 Induced expression of IL-6 ( 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 increased expression of Dicer decreased response to H 2 o 2 Sensitivity to induced DNA damage ( Figure 14 B-D and Figure 15 B-D). Thus, treatment with anastrozole reduces H 2 o 2 Induced increased expression of cytoplasmic DNA and IL-6 ( Figure 14 E-F and Figure 16 A-B). Silencing Dicer expression could partially eliminate the effect of anastrozole ( Figure 17 A-C and Figure 18 A-B). Together, these findings suggest that anastrozole...
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