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A biomarker related to the occurrence and development of breast cancer and its application

A kind of breast cancer and material technology, applied in the field of biomedicine, can solve the problem of insufficient breast cancer

Active Publication Date: 2020-06-02
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the diversity of breast cancer genetic background and response to treatment, these indicators for evaluating breast cancer are far from enough. We need more and better biomarkers to guide early diagnosis, choose appropriate treatment methods and accurately predict Prognosis

Method used

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  • A biomarker related to the occurrence and development of breast cancer and its application
  • A biomarker related to the occurrence and development of breast cancer and its application
  • A biomarker related to the occurrence and development of breast cancer and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] Example 1 Screening of gene markers related to breast cancer

[0076] 1. Sample collection

[0077] Collect paracancerous tissues and breast cancer tissue samples from 8 breast cancer patients with a clear pathological diagnosis, and state the sample name, tissue type, number, sampling date, sample processing process, etc., and all patients signed an informed consent. All were approved by the organization ethics committee.

[0078] 2. Preparation of RNA samples

[0079] Use QIAGEN's tissue RNA extraction kit to extract RNA samples. See the instructions for specific operations.

[0080] 3. Reverse transcription and labeling

[0081] The mRNA was reverse transcribed into cDNA with Low RNA Input Linear Amplification Kit, and the experimental group and control group were labeled with Cy3.

[0082] 4. Hybridization

[0083] The gene chip uses Kangcheng Bio-Human lncRNAArray, and hybridizes according to the steps in the chip instruction manual.

[0084] 5. Data analysis

[0085] Use Agilen...

Embodiment 2

[0088] Example 2 QPCR sequencing to verify the differential expression of LOC100996251 gene

[0089] 1. Perform large sample QPCR verification on the differential expression of LOC100996251 gene. According to the sample collection method in Example 1, 50 cases of breast cancer adjacent tissue and 50 cases of breast cancer tissue were selected.

[0090] 2. RNA extraction

[0091] Use QIAGEN's tissue RNA extraction kit to extract RNA samples. See the instructions for specific operations.

[0092] 3. QPCR

[0093] 1) Reaction system:

[0094] RNA template 1μl, random primer 1μl, double distilled water to 12μl, mix well, centrifuge at low speed, 65°C for 5min, then cool on ice.

[0095] Continue to add the following ingredients to the 12μl reaction solution:

[0096] 5× reaction buffer 4μl, RNase inhibitor (20U / μl) 1μl, 10mM dNTP mixture 2μl, AMV reverse transcriptase (200U / μl) 1μl; mix well and centrifuge;

[0097] 2) Reverse transcription reaction conditions

[0098] 25℃ for 5min, 42℃ for 60m...

Embodiment 3

[0117] Example 3 Expression of LOC100996251 in breast cancer cell lines

[0118] 1. Cell culture

[0119] Cultivation of human breast cancer cell lines MCF-7, SK-BR-3, MDA-MB-231 and normal breast epithelial cell line MCF-l0A, of which MDA-MB-231 was cultured in L15 medium with 10% fetal bovine serum , SKBR3 was cultured in RPMI-1640 medium containing 10% fetal bovine serum, MCF-7 and normal breast epithelial cell line MCF-10A were cultured in DMEM medium containing 10% fetal bovine serum. Add 1% P / S to the culture solution. At 37℃, 5% CO 2 , Cultivation in an incubator with a relative humidity of 90%. Change the medium once every 2-3 days, and use 0.25% trypsin containing EDTA for routine digestion and passage.

[0120] 2. RNA extraction and concentration determination

[0121] Use QIAGEN's Cell RNA Extraction Kit to extract total cell RNA. Refer to the instructions for specific operations.

[0122] 3. The specific steps of QPCR are the same as in Example 2

[0123] 4. Statistical a...

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Abstract

The invention discloses a biomarker relevant to breast cancer occurrence and development and application thereof. The biomarker is LOC100996251. The invention discloses a reagent, a kit and a chip fordetecting the level of the LOC100996251. The invention also discloses a composition containing an LOC100996251 accelerant.

Description

Technical field [0001] The present invention belongs to the field of biomedicine and relates to a biomarker related to the occurrence and development of breast cancer and its application. Specifically, the lncRNA is LOC100996251. Background technique [0002] Breast cancer is one of the most common malignant tumors in women worldwide (DeSantis CE, Fedewa SA, Goding Sauer A, Kramer JL, Smith RA, Jemal A. Breast cancer statistics, 2015. CACancer J Clin. 2016; 66(1) :31--42.). The occurrence of breast cancer is accompanied by changes in genes, epigenetics and various molecular mechanisms (Manca G, Tardelli E, Rubello D, et al. Nucl MedCommun. 2016; 37(6): 574-576.). With the advancement of early diagnosis of breast cancer, as well as the development of radiotherapy and various adjuvant treatments, the prognosis of breast cancer has been greatly improved, but breast cancer is still the most common malignant tumor that threatens the lives of women (Fan L , Goss PE, Strasser-Weippl K...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886A61K31/7088A61P35/00
CPCA61K31/7088C12Q1/6886C12Q2600/136C12Q2600/158C12Q2600/178
Inventor 孙锦云石小峰
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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