Application of noncoding gene in diagnosis and treatment of liver cancer

A liver cancer and gene technology, applied in the field of non-coding protein gene LINC01703

Active Publication Date: 2018-01-16
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current mechanism of action of molecular targeted drugs is only limited to the superficial curative effect, and the deep-seated mechanism still needs further research

Method used

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  • Application of noncoding gene in diagnosis and treatment of liver cancer
  • Application of noncoding gene in diagnosis and treatment of liver cancer
  • Application of noncoding gene in diagnosis and treatment of liver cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Example 1 Screening for gene markers associated with liver cancer

[0073] 1. Sample collection

[0074] The cancer tissues and paracancerous tissues of 10 patients with liver cancer were collected, and the patients gave informed consent. All the above specimens were obtained with the consent of the organizational ethics committee.

[0075] 2. Preparation of RNA samples

[0076] Tissue RNA was extracted using QIAGEN tissue RNA extraction kit, and the operation was performed according to the specific steps in the manual.

[0077] 3. Reverse transcription and labeling

[0078] The mRNA was reverse-transcribed into cDNA using the Low RNA Input Linear Amplification Kit, and the experimental group and the control group were labeled with Cy3, respectively.

[0079] 4. Hybridization

[0080] The gene chip uses Cannes Bio-Human lncRNA Array, and hybridization is performed according to the steps of the chip instruction manual.

[0081] 5. Data processing

[0082] After hybri...

Embodiment 2

[0085] Example 2 QPCR sequencing to verify the differential expression of LINC01703 gene

[0086] 1. Large-scale QPCR verification of the differential expression of the LINC01703 gene. According to the sample collection method in Example 1, 60 samples of liver cancer tissues and 60 samples of paracancerous tissues were collected.

[0087] 2. The RNA extraction steps are the same as in Example 1.

[0088] 3. Reverse transcription:

[0089] Use 25μl reaction system, take 1μg total RNA for each sample as template RNA, and add the following components to PCR tubes: DEPC water, 5× reverse transcription buffer, 10mM dNTP, 0.1mM DTT, 30μM Oligo dT, 200U / μl M-MLV, template RNA. Incubate at 42°C for 1h, then centrifuge briefly at 72°C for 10min.

[0090] (3) QPCR amplification test

[0091] The amplification primer sequences of LINC01703 and housekeeping gene GAPDH were designed and synthesized by Shanghai Sangong. The primer sequences of LINC01703 gene are shown in SEQ ID NO.2~3;...

Embodiment 3

[0097] Example 3 Analysis of the expression of LINC01703 in the TCGA database

[0098] 1. Data collection

[0099] The lncRNA expression profile data of 200 cases of liver cancer tissues and 50 cases of para-cancerous tissues were collected from the TCGA database, and the expression level of LINC01703 in liver cancer tissues and para-cancerous tissues was analyzed; box plots were drawn.

[0100] 2. ROC curve analysis

[0101] The receiver operating characteristics of LINC01703 were analyzed using the pROC package in R language, the binomial exact confidence space was calculated, and the ROC curve was drawn.

[0102] 3. Results

[0103] The expression level of LINC01703 as figure 2 As shown, compared with the control group, the expression of LINC01703 was significantly up-regulated in liver cancer tissues.

[0104] The ROC curve of LINC01703 is as image 3 As shown, the AUC value of LINC01703 is as high as 0.905, and has high specificity and sensitivity, indicating that L...

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Abstract

The invention discloses application of a noncoding protein gene in the diagnosis and treatment of liver cancer, and the specific noncoding protein gene is LINC01703. The invention discovers for the first time that LINC01703 is related to the incidence and development of liver cancer. By bioinformatic analysis, the invention discovers that LINC01703 has relatively high AUC value in patients with liver cancer, indicating that LINC01703 has relatively high accuracy and specificity as a diagnostic marker for liver cancer. Through an experiment, the invention verifies that changing the expression level of LINC01703 can influence the proliferation and apoptosis and the like of liver cancer, indicating that LINC01703 can be used as a drug target in the treatment of liver cancer.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to the application of a non-coding protein gene in the diagnosis and treatment of liver cancer, in particular to the non-coding protein gene LINC01703. Background technique [0002] Hepatocellular carcinoma (HCC) is the most common malignant tumor worldwide, especially in Asia, Africa and southern Europe. HCC has a high degree of malignancy and a short survival period, with a 5-year survival rate of about 3% to 5%. Due to the rich blood supply and lymphatic circulation of the liver, cancer cells are prone to spread and metastasis, so the mortality rate of advanced liver cancer is extremely high. my country's annual new cases account for about 45% of the world, and the annual mortality rate of malignant tumors ranks first and second in rural and urban malignant tumor mortality, and has become the country with the highest incidence of liver cancer. The occurrence of liver cancer seriously en...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/11A61K48/00A61K45/00A61K31/713A61P35/00
Inventor 宋宏涛马翠任静
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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