Application of substance for inhibiting expression of ZNF8 protein in preparing products for preventing and treating cancer

A technology of expression quantity and product, applied in the field of biomedicine

Pending Publication Date: 2019-05-28
BEIJING PROTEOME RES CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Its function and mechanism of a

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  • Application of substance for inhibiting expression of ZNF8 protein in preparing products for preventing and treating cancer
  • Application of substance for inhibiting expression of ZNF8 protein in preparing products for preventing and treating cancer
  • Application of substance for inhibiting expression of ZNF8 protein in preparing products for preventing and treating cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0086] Embodiment 1, the construction of plasmid, the preparation of oligonucleotide and the acquisition of cell

[0087] 1. Construction of Myc-ZNF8 plasmid

[0088] The small fragment between the recognition sequences of the restriction endonucleases EcoRI and XhoI of the pCMV-Myc plasmid is replaced with the DNA molecule shown in sequence 2 in the sequence listing, and the obtained recombinant plasmid is the Myc-ZNF8 plasmid.

[0089] The Myc-ZNF8 plasmid expresses the ZNF8 protein shown in Sequence 1 in the Sequence Listing.

[0090] 2. Construction of Flag-p53 plasmid

[0091] The small fragment between the recognition sequences of the restriction endonucleases EcoRI and BamHI of the pCMV-Flag plasmid is replaced with the DNA molecule shown in sequence 4 in the sequence table, and the resulting recombinant plasmid is the Flag-p53 plasmid.

[0092] The Flag-p53 plasmid expresses the p53 protein shown in sequence 3 in the sequence listing.

[0093] 3. Preparation of olig...

Embodiment 2

[0119] Embodiment 2, suppress the expression of ZNF8 protein and promote p53 protein activity

[0120] Experiment 1. Inhibiting the expression of ZNF8 protein promotes the transcriptional activity of endogenous p53 protein

[0121] The experiment was repeated three times to take the average value, and the steps for each repetition were as follows:

[0122] 1. Cells (p53 + / + HCT116 cells or p53 - / - HCT116 cells) in 6 wells of a 24-well plate containing 0.5mL DMEM medium (8.0×10 per well 4 cells), and then placed at 37°C, 5% CO 2 Culture in an incubator, and when the fusion rate reaches 70-90%, they are randomly divided into two groups, and each group is set with three replicate wells, and the following treatments are carried out:

[0123] The first group: add 20ng pG13L plasmid, 0.2ng pRL-TK plasmid and 0.4μg shNC plasmid to each well, and co-transfect for 36h.

[0124] The second group: add 20ng pG13L plasmid, 0.2ng pRL-TK plasmid and 0.4μg shZNF8-1 plasmid to each well, ...

Embodiment 3

[0139] Example 3, ZNF8 protein and p53 protein all interact under normal conditions and DNA damage conditions

[0140] Co-immunoprecipitation experiments were used to detect the interaction between ZNF8 protein and p53 protein under normal conditions and DNA damage conditions.

[0141] Experiment 1. Under normal conditions, endogenously expressed ZNF8 protein can interact with p53 protein

[0142] The experiment was repeated three times, and the steps for each repetition were as follows:

[0143] 1. Convert p53 + / + HCT116 cells were inoculated in a culture bottle containing 30 mL of DMEM medium (the size of the culture bottle was 25 cm 2 ;3.0×10 per flask 6 cells) at 37°C, 5% CO 2 Cultivate in the incubator for 48 hours (at this point the fusion rate reaches 100%).

[0144] 2. After completing step 1, add trypsin digestion solution (for the purpose of digesting the cells) into the culture flask, then transfer to a centrifuge tube (50mL), centrifuge at 1000rpm for 5min, an...

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Abstract

The invention discloses application of a substance for inhibiting the expression of ZNF8 protein in preparing products for preventing and treating cancer. An experiment shows that the ZNF8 protein canbe bound with p53 protein; by inhibiting the expression of the ZNF8 protein, the transcription activity of the p53 protein can be increased, the expressions of a VEGF gene and an ANGPLT4 gene are reduced, and proliferation, metastasis, migration, invasion and metastasis of tumor cells are inhibited; the formation of blood vessels is inhibited; the growth,the metastasis and lung metastasis of thetumor are inhibited; the survival rate of tumor-carrying animals is improved. Therefore, inhibition of the expression of the ZNF8 protein has important application value in inhibiting the growth and invasive metastasis of the tumor. Cancer can be prevented and/treated by inhibiting the expression of the ZNF8 protein.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to the application of a substance inhibiting the expression of ZNF8 protein in the preparation of products for preventing and treating cancer. Background technique [0002] The KRAB zinc finger protein (KRAB-containing zinc finger protein, KZNF) family is the largest transcription factor / transcription regulator family in mammals, and its protein structure is mainly composed of the conserved KRAB (Krüppel -associated box) domain, the middle and the C-terminus contain multiple C2H2-type zinc finger structures that continuously recognize continuous DNA sequences. Among them, the KRAB domain recruits a variety of transcriptional repressors by combining with KAP-1 (KRAB-associated protein-1), forming a transcriptional repression complex, and the DNA and / or other transcriptional factors in the C-terminal zinc finger region and the target gene regulatory region After binding, the transcriptiona...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K45/00A61K31/7105A61P35/00A61P35/04G01N33/68
Inventor 田春艳王建张秀园贺福初
Owner BEIJING PROTEOME RES CENT
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