Application of p53 gene mutation in cancer-associated fibroblasts

A fibroblast, tumor-related technology, applied in the determination/inspection of microorganisms, instruments, measuring devices, etc., can solve the problems of drug resistance hindering treatment, and the regulatory effect has not yet been reported.

Pending Publication Date: 2018-09-04
KUNMING UNIV OF SCI & TECH
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0003] At present, tumor treatment mainly includes surgery, chemotherapy, radiotherapy, immunotherapy, hormone therapy and targeted gene therapy; patients can obtain obvious effects in the early stage of treatment, but the emergence of drug resistance seriously hinders subsequent treatment; after a long time, tumor The treatment and drug resistance of tumors are mostly concentrated in the tumor itself, ignoring the influence of CAFs on the tumor; developing reliable markers to determine the activation and characteristics of CAFs, so as to formulate personalized treatment plans and adjust the tumor microenvironment, can effectively improve the Cancer therapy effect; the CAF characteristics of p53S-activated cells suggest that it may be a good therapeutic target. At present, the regulatory effect of p53S on the tumor microenvironment in non-tumor stromal cells has not been reported

Method used

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  • Application of p53 gene mutation in cancer-associated fibroblasts
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  • Application of p53 gene mutation in cancer-associated fibroblasts

Examples

Experimental program
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Embodiment 1

[0011] Example 1: Preparation of p53S MEFs

[0012] 1. Observation of vaginal suppositories

[0013] to get more p53 S / S MEF, we choose p53 S / S (♂)× p53 S / + (♀) for hybridization; put one male and one male mouse of the right age together in the afternoon (usually 2-4 month old mice), and observe whether there is a vaginal suppository in the vagina of the female mouse before 9:00 the next morning, If a vaginal suppository is seen, the male and female should be kept separately, and the day when the vaginal suppository is seen is recorded as 0.5 days.

[0014] 2. Preparation of MEFs

[0015] The 13.5-day-pregnant female mouse was killed by neck dislocation, and put into alcohol with a mass concentration of 70%; after dissection, the embryos in the mother were taken out and placed in a plate with 1 × PBS, and one of the embryos was cut out and placed in another containing 1 ×PBS plate, remove the maternal uterine tissue, outer membrane and heart, rinse in 1×PBS and put in...

Embodiment 2

[0049] Embodiment 2: Construction of mouse xenograft tumor model

[0050] 1. Prepare male nude mice aged 6-8 weeks, according to the weight of the mice, and refer to the sex of the mice (the tumor cells used in this experiment are PC-3 prostate cancer cells, and the fibroblasts are male WT, P53 S / S cells), mice were randomly divided into 3 groups (PC-3, mixed PC-3+WT, mixed PC-3+ p53 S / S ), with 3-5 mice per group;

[0051] 2. Melt BD Matrigel at 4°C one day in advance;

[0052] 3. Take the P4 generation cells that grow to about 90%, digest with 0.01% trypsin for 1-2min, stop the digestion with a small amount of medium, and collect the cell suspension in a sterile 15mL centrifuge tube;

[0053] 4. Mix the cell suspension, take 25 μL of the cell suspension, add it to 100 μL 1XPBS (5-fold dilution), mix well, and count with a hemocytometer; (The amount of cells at each injection site of PC-3 cells is 2 ×10 6 each / 50μL, PC-3 and MEF (WT, p53 S / S ) Mix according to the rati...

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Abstract

The invention discloses application of p53 gene mutation in cancer-associated fibroblasts, and particularly relates to application of p53 gene mutation in the cancer-associated fibroblasts in screening and preparation of cancer therapy medicines. An experimental result shows that the growth of cancer can be promoted by mouse embryonic fibroblasts (MEF) carrying p53N236S mutation, thus it is suggested that p53 mutation activates the cancer-associated fibroblast characteristic of the MEF, and the favorable microenvironment is provided for multiplication, migration and the like of the cancer. Thenew function of p53 mutation in the cancer-associated fibroblasts is disclosed for the first time, through the discovery, the new idea and research direction are provided for p53 mutation in cancer research, a new action target is provided for cancer therapy, and a new strategy is also provided for cancer therapy.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the application of p53 gene mutation in tumor-associated fibroblasts in screening and preparing tumor therapeutic drugs. Background technique [0002] Tumor is a major disease that seriously endangers human health. Its occurrence and development are a complex process; it not only involves cancer cells that drive the malignant process of tumors, but also includes various non-tumor stromal cells that constitute the tumor microenvironment. Among them, tumor-associated fibroblasts (cancer-associated fibroblasts, CAFs) are an important part of the tumor microenvironment. CAFs secrete chemokines, cytokines, inflammatory factors, etc., provide a favorable microenvironment for tumors, promote tumor growth, metastasis, and induce angiogenesis. [0003] At present, tumor treatment mainly includes surgery, chemotherapy, radiotherapy, immunotherapy, hormone therapy and targeted gene...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/02
CPCC12Q1/6886C12Q2600/156G01N33/5091
Inventor 吴晓明田营彬罗瑛
Owner KUNMING UNIV OF SCI & TECH
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