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Molecular marker COL11A1 and application thereof to breast cancer drugs

A molecular marker, breast cancer technology, applied in the field of oncology and molecular biology, can solve the problems of thinning collagen fibers, reducing tissue tensile strength, affecting tumor invasion and migration, etc.

Pending Publication Date: 2021-03-16
NORTHWESTERN POLYTECHNICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The expression of COL11A1 in stromal cells around the tumor tissue increases, resulting in thinning of collagen fibers and reducing the tensile strength of the tissue. Therefore, COL11A1 may participate in the epithelial-mesenchymal transition (Epithelial-Mesenchymal Transition, EMT) pathway, thereby affecting tumor growth and development. invasion and migration

Method used

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  • Molecular marker COL11A1 and application thereof to breast cancer drugs
  • Molecular marker COL11A1 and application thereof to breast cancer drugs
  • Molecular marker COL11A1 and application thereof to breast cancer drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1 Discovery of COL11A1 as a breast cancer marker.

[0042]1. Set the search scope of the Oncomine database to "Analysis Type>Cancer vs. NormalAnalysis>Cancer Type>Breast Cancer>Data Type>mRNA>Molecular Subtype>Triple Negative Status", so as to extract the breast cancer sample information and analyze the data. Statistics finishing. Afterwards, screening was performed according to the expression difference and p-value of genes in different samples. The selected genes were searched in the web of science database to investigate their reliability as markers and their functions in tumors. The gene COL11A1 was finally identified.

[0043] 2. Extraction of total RNA from breast cancer tissue and cells

[0044] a. For tissue: Take an appropriate amount of tumor tissue from a breast cancer patient and place it in a 1.5mL non-enzyme centrifuge tube and cut it into pieces as small as possible with non-enzyme surgical scissors, then add 1mL of TRIzol lysis solution, and g...

Embodiment 2

[0076] Example 2 Detection of the effect of COL11A1 on the function of MCF-7 in breast cancer cells

[0077] 1. Design of COL11A1-siRNA

[0078] According to published literature reports and the recommendation of the siRNA design website (https: / / www.thermofisher.com / ), five siRNAs of COL11A1 gene were preliminarily screened, and the specific sequences are shown in Table 5. siRNA was synthesized by Shanghai Gema Pharmaceutical Technology Co., Ltd.

[0079] Table 5 siRNA sequences of COL11A1 gene

[0080]

[0081]

[0082] 2. 24h after transfection of siRNA into MCF-7 cells, the cells were harvested, and total cell RNA was extracted. After reverse transcription, the siRNA knockdown efficiency was detected by qPCR.

[0083] 3. Detect the effect of COL11A1-siRNA on MCF-7 cell cycle

[0084] a. MCF-7 cells in good growth condition were divided into 1 × 10 6 The amount per well was inoculated into a 6-well plate, and COL11A1-siRNA was transfected after the growth state wa...

Embodiment 3

[0108] Example 3 Effect of COL11A1-siRNA drug on tumor in animals

[0109] 1. Construction of COL11A1-siRNA local sustained release system

[0110] The local sustained-release system of miR-4458 was constructed with gelatin nanospheres (GNs) as the carrier, and the purpose of long-acting sustained-release was achieved after in situ injection. The specific construction steps are as follows:

[0111] a. Weigh 3.2 mg of GNs, sterilize them with UV light for 30 min, and transfer them to a sterile RNase free centrifuge tube;

[0112] b. Add 6.5 μL 2000 was added to 9uL OPTI-MEM, 2μL siRNA was added to 6.5μL OPTI-MEM, gently mixed, and allowed to stand at room temperature for 5min;

[0113] c. will contain Add 2000 OPTI-MEM mixture to siRNA-containing OPTI-MEM mixture, mix gently, and let stand at room temperature for 20 minutes;

[0114] d. will The 2000-siRNA-OPTI-MEM mixture was added to a sterile RNasefree centrifuge tube containing GNs, and allowed to stand at 4°C for ...

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Abstract

The invention discloses a molecular marker COL11A1 and an application thereof to breast cancer drugs, and belongs to the field of oncology and molecular biology. The Gene ID of the gene sequence of the COL11A1 is 1301, and the expression of the COL11A1 in a breast cancer biological sample is up-regulated. Through Oncomine database collection and screening and patient tissue sample and breast cancer cell inspection based on a qRT-PCR technology, it is proved that the COL11A1 is significantly up-regulated in breast cancer tissues and cells. The invention proves the reliability and the repeatability of the COL11A1 as the breast cancer molecular marker. It is proved that proliferation, clone formation, migration and invasion of breast cancer cells MCF-7 are remarkably inhibited by knocking down expression of the COL11A1 through siRNA. Therefore, the breast cancer can be accurately diagnosed by using the COL11A1 as the marker, and a therapeutic target and an important theoretical basis areprovided for gene therapy, small molecular drugs and other clinical applications.

Description

technical field [0001] The invention belongs to the field of oncology and molecular biology, and particularly relates to a molecular marker COL11A1 and its application in breast cancer medicine. Background technique [0002] Breast cancer ranks first in the incidence of female malignant tumors in my country, and the mortality rate ranks fifth. Almost all breast cancer-related deaths are caused by metastasis. Endocrine therapy is the standard and effective therapy for breast cancer. Endocrine therapy can make breast cancer patients recur. The relative risk of breast cancer is reduced by about 40%, which improves the prognosis of breast cancer patients and improves the quality of life of patients. However, due to changes in the breast cancer cells themselves and their surrounding microenvironment, about 1 / 3 of patients develop drug resistance and recurrence after long-term endocrine drug treatment, so it is very important to find new treatment methods. [0003] The extracellul...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/158
Inventor 张辰艳刘杰杨长青赵士淇张葛张世龙罗力恒尹大川
Owner NORTHWESTERN POLYTECHNICAL UNIV
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