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Breast tumor prognosis biomarker LncRNA detection method and clinical application thereof

A breast tumor and detection method technology, applied in the field of breast tumor prognostic biomarker LncRNA detection, can solve problems such as breast tumor diagnosis and gene therapy impact, and achieve the effect of guiding prognosis clinical treatment, improving prognosis effect evaluation, and low detection cost.

Inactive Publication Date: 2016-01-13
杭州壹锋生物科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These findings all indicate that LncRNA plays an important role in cell proliferation, differentiation and carcinogenesis, and the discovery of breast tumor-associated LncRNA may have a significant impact on the diagnosis and gene therapy of breast tumors

Method used

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  • Breast tumor prognosis biomarker LncRNA detection method and clinical application thereof
  • Breast tumor prognosis biomarker LncRNA detection method and clinical application thereof
  • Breast tumor prognosis biomarker LncRNA detection method and clinical application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1. Solexa sequencing technology performs high-throughput sequencing of long non-coding RNAs in cancer tissues of breast cancer patients.

[0053] 1. Extraction of total RNA from breast cancer and corresponding paracancerous tissues

[0054] Tissue Lysis:

[0055] 1) In a clean area with less RNase interference (all utensils are wiped with 1‰ DEPC water), the mortar needs to be sterilized at 200°C for 5 hours and cooled for 4 hours. In a mortar pre-cooled with liquid nitrogen, grind to powder with a pestle (incomplete grinding will seriously affect the yield of RNA);

[0056] 2) Add 700 μl of QIAzol lysate to the mortar, continue to grind to a homogeneous liquid without obvious tissue pieces, and then transfer to a RNase-free 1.5mleppendorf tube;

[0057] 3) Refrigerated centrifuge at 4°C, 12000×g, and centrifuge for 10 minutes to remove incompletely lysed tissues and cell debris in the solution;

[0058] 4) Carefully absorb the supernatant after centrifugatio...

Embodiment 2

[0117] Example 2 According to the sequencing results obtained in solexa, the obtained data were screened for LncRNA and analyzed for different pathologies.

[0118] Quantitative analysis of lncRNA expression and its expression correlation in different pathological indicators

[0119] The information of breast cancer tissue and adjacent tissue samples is according to the database website http: / / www.cbioportal.org / , and for breast cancer, press "Mutations Putative copy-number alterations from GISTIC mRNAExpressionz-Scores (RNASeqV2RSEM)" was screened from 959 breast samples for HUGOgenenomenclaturecommittee( http: / / www.genenames.org / ) for the analysis of 2,731 lncRNAs recognized in ), and 959 breast patient samples with complete pathological information were screened out. The information of 959 cases is as follows:

[0120] Table 1 Clinicopathological information of patients with breast cancer and adjacent tissues

[0121]

[0122]

[0123] (1) The expression of lnc...

Embodiment 3

[0137] Example 3 Linc00657 expression difference analysis between breast cancer tissue and normal tissue ( Figure 8 )

[0138] In this experiment, OriGenebreastcancercDNAarrays were used to conduct RT-PCR experiments, and the experimental data obtained by RT-PCR were analyzed to obtain the relative change multiples in breast cancer tissue and normal tissue samples, and then use the independent sample T test in SPSS statistical analysis software to analyze The expression of Linc00657 in 41 cases of breast cancer tissue samples and 7 cases of normal tissue samples in the experiment, the expression level of Linc00657 in breast cancer tissues was 1.95 times higher than that in normal tissues, and it was statistically significant (P=0.01). This experiment further verified the sequencing experiment results of 959 breast patients in TCGA database analysis.

[0139] (1) Detection of primer combinations. Invitrogen Beijing Branch was commissioned to synthesize the following primers fo...

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Abstract

The invention discloses a breast tumor prognosis biomarker LncRNA detection method and clinical application thereof, provides a group of new LncRNA in sequence shown as SEQ ID No1, SEQ ID No2, SEQ ID No3, SEQ ID No4 and SEQ ID No5 and relates to a method of taking specific LncRNA in samples (tissues, serum, urine, body fluid and the like) of patients suffering from breast tumors and corresponding juxtacancerous samples or normal samples as detection markers and biomarkers for prognostic auxiliary detection of breast tumors, related kits and a high-throughput sequencing and screening method. In-vitro diagnosis and judgment of canceration and process of breast tumors are realized by identification of differences of LncRNA expression quantity in the breast tumor samples and the corresponding juxtacancerous samples or normal samples. The invention further provides usage of the LncRNA as a breast tumor marker and probes and primers for LncRNA detection.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a method and clinical application for detecting LncRNA, a prognostic biomarker of breast tumors. Background technique [0002] There are about 20,000 to 30,000 genes that encode proteins in the human body, accounting for only 2% of the human genome, and the remaining 98% of genomic DNA that does not encode proteins was originally considered to be nonfunctional and garbage in organisms, often called "Junk DNA. However, current research shows that most of these junk DNA can be transcribed to produce non-coding RNA (non-codingRNA, ncRNA). According to the size of mature transcripts, ncRNA can be divided into small molecule ncRNA (such as siRNA, miRNA, piRNA etc.), medium-length ncRNA (70-200nt) and long ncRNA (longncRNA, LncRNA, >200nt). At present, the research on ncRNA is mostly small molecule ncRNA, and the research on LncRNA is still in its infancy. Due to the presence ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12Q1/68C12N15/11
Inventor 丁先锋莫寅元
Owner 杭州壹锋生物科技有限公司
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