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LncRNA related to colorectal cancer and application thereof

A colorectal cancer and reagent technology, applied to lncRNA and its application in the preparation of disease diagnostic reagents, can solve the problems of abnormal expression of 1ncRNAs, small size, and difficulty in finding, etc.

Pending Publication Date: 2019-05-10
TIANJIN CITY THIRD CENT HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In terms of diagnosis, since the expression of 1ncRNAs in tumor tissue is often abnormal and has tissue and species specificity, it may become a tumor marker, especially in the early stage of some tumors. Due to its small size, traditional imaging methods are not easy Find

Method used

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  • LncRNA related to colorectal cancer and application thereof
  • LncRNA related to colorectal cancer and application thereof
  • LncRNA related to colorectal cancer and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Sample Collection Extraction and Data Analysis

[0034] Collect cancer and paracancerous tissues from 4 patients undergoing surgery for rectal adenocarcinoma in our hospital, put them into 1.5ml sterile EP tubes soaked in 0.1% DEPC water overnight and sterilize, and quickly put them into a liquid nitrogen tank at -80°C Save within. After extracting sample RNA, construct lncRNA and mRNA libraries, and then use llumina hiseq x-ten second-generation high-throughput sequencing technology to sequence mRNA. The data processing is completed through processes such as joint removal, low quality removal, and decontamination to obtain sequencing data.

[0035] Quantification of mRNA expression and differential expression analysis were performed using software cuffquan and cuffdiff. cuffquant is a tool specially used in the Cufflinks suite for expression evaluation and standardization. cuffdiff is a tool used in the Cufflinks suite to calculate differential expression. ...

Embodiment 2

[0036] Co-expression gene analysis of embodiment 2LOC105374879

[0037] As a kind of non-coding RNA, lncRNA's function is mainly reflected in the regulation of target genes, mainly including the trans-regulation of distant protein-coding genes. At the same time, genes with the same expression pattern, in the There is a strong correlation in function. Therefore, the target genes of lncRNA were studied by co-expression and trans analysis of lncRNA and mRNA.

[0038] The co-expression relationship between lncRNA and mRNA was analyzed by calculating the Pearson correlation coefficient (Pearson correlation coefficient, PCC) of differentially expressed lncRNA and mRNA expression. The threshold value was 0.98, and pfigure 1 ), HSPA5, TDP1, COA7, C1QBP, ASNS, CCNE1, HEATR1, TMEM201, ADK, RANBP1, GUCA2A, CLDN23, CD79A, RAB27A, OTUD1, TMEM54, ABHD3, RIMKLA, USP2, CD244, among which the positively correlated genes are: HSPA5 , TDP1, COA7, C1QBP, ASNS, CCNE1, HEATR1, TMEM201, ADK, RANBP1...

Embodiment 3

[0039] Example 3 Fluorescent quantitative PCR verification of differentially expressed lncRNA

[0040] 1. Sample collection and extraction

[0041] For sample collection, 28 cases of peripheral blood (including 13 cases of colon cancer and 15 cases of rectal cancer) diagnosed as colon cancer or rectal cancer in our hospital were selected, and 24 cases of peripheral blood of healthy people who underwent physical examination were selected as controls. Blood RNA was extracted by QIAGEN The kit extracts RNA from serum, and the specific steps refer to the instructions.

[0042] 2. Reverse transcription:

[0043] 1) Mix 10pg-1μg total RNA template with 2μl 10× buffer, 2μl dATP (10mM), 0.5μl polyA polymerase, 0.5μl RNase inhibitor and RNase free water , the final volume was 20 μl, and incubated at 37°C for 1h.

[0044] 2) Add 1 μl of 0.5 μg / μl Oligo(dT) specific RT primer to the reaction tube, and incubate at 70° C. for 5 minutes.

[0045] 3) Immediately incubate on ice for at le...

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Abstract

The invention discloses an lncRNA related to colorectal cancer and application thereof. An inventor conducts transcriptome lncRNA-mRNA sequencing analysis on cancer tissue and para-carcinoma tissue ofpatients suffering from the colorectal cancer, 1ncRNAs obvious in differential expression and co-expression genes thereof are sought, and LOC105374879 closely related to the colorectal cancer and co-expression genes of the LOC105374879 are found. Accordingly, a new molecular marker is provided for the early diagnosis of the colorectal cancer, and a foundation is laid for clinical application.

Description

technical field [0001] The invention belongs to the field of cancer molecular diagnosis, and in particular relates to an lncRNA related to colorectal cancer and its application in the preparation of disease diagnostic reagents. Background technique [0002] Colorectal cancer (colorectal, CRC) is one of the most common malignant tumors, the domestic incidence rate has been increasing in recent years, and has become one of the nine common tumors in my country. In recent decades, the improvement of various detection methods has made great progress in the diagnosis of colorectal cancer. Currently, colorectal cancer screening in my country mainly adopts the sequential screening method. That is, the high-risk factor questionnaire combined with the immunological method of fecal occult blood test (FOB) is used as the initial screening program. People who are positive in the questionnaire or FOB are considered to be positive in the initial screening and will undergo colonoscopy. Th...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886
Inventor 李成龙张自立吴迎梅张金卷朱争艳张艳艳李南南景丽刘彤王鹏
Owner TIANJIN CITY THIRD CENT HOSPITAL
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