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Gene related to liver cancer and application thereof

A liver cancer, gene technology, applied in ENSG00000260285 gene and its application field, can solve the problem of lack of systematic research and so on

Active Publication Date: 2017-10-03
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The role of lncRNA in HCC is still unknown, and there is a lack of systematic research. It is of great significance to reveal the function and mechanism of lncRNA in hepatocellular carcinoma

Method used

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  • Gene related to liver cancer and application thereof
  • Gene related to liver cancer and application thereof
  • Gene related to liver cancer and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] Example 1 Screening Gene Markers Related to Liver Cancer

[0075] 1. Sample collection

[0076] 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.

[0077] 2. Preparation of RNA samples

[0078] Tissue RNA was extracted using QIAGEN tissue RNA extraction kit, and the operation was performed according to the specific steps in the manual.

[0079] 3. Reverse transcription and labeling

[0080] 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.

[0081] 4. Hybridization

[0082] The gene chip uses Cannes Bio-Human lncRNA Array, and hybridization is performed according to the steps of the chip instruction manual.

[0083] 5. Data processing

[0084] After hybridizatio...

Embodiment 2

[0087] Example 2 QPCR sequencing to verify the differential expression of the ENSG00000260285 gene

[0088] 1. Large sample QPCR verification of differential expression of ENSG00000260285 gene. According to the sample collection method in Example 1, 60 samples of liver cancer tissues and 60 samples of paracancerous tissues were collected.

[0089] 2. RNA extraction The steps are the same as in Example 1.

[0090] 3. Reverse transcription:

[0091] 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 1 hour, then centrifuge briefly at 72°C for 10 minutes.

[0092] (3) QPCR amplification test

[0093] Primer design:

[0094] The primer sequence of ENSG00000260285 gene is:

[0095] Forward primer: 5'-TTCTGCCTGGACAACATAG-3' (SEQ ID NO.2)

[0096] Reverse primer: 5'-AATCAGC...

Embodiment 3

[0105] Example 3 Analysis of the expression of ENSG00000260285 in the TCGA database

[0106] 1. Data collection

[0107] The lncRNA expression profile data of 200 cases of liver cancer tissues and 50 cases of paracancerous tissues were collected from the TCGA database, and the expression level of ENSG00000260285 in liver cancer tissues and paracancerous tissues was analyzed; box plots were drawn.

[0108] 2. ROC curve analysis

[0109] The receiver operating characteristics of ENSG00000260285 were analyzed using the pROC package in R language, the binomial exact confidence space was calculated, and the ROC curve was drawn.

[0110] 3. Results

[0111] The expression level of ENSG00000260285 as figure 2 As shown, compared with the control group, the expression of ENSG00000260285 was significantly up-regulated in liver cancer tissues.

[0112] The ROC curve of ENSG00000260285 is as follows image 3 As shown, the AUC value of ENSG00000260285 is as high as 0.938, and has hig...

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Abstract

The invention discloses a gene related to liver cancer and application thereof. Specifically, the gene is ENSG00000260285. According to the gene disclosed by the invention, up-regulated expression of the ENSG00000260285 in a liver cancer patient is found for the first time through high-throughput sequencing, and differential expression of the gene in the liver cancer patient is further verified by qPCR. Bioinformatic analysis finds that the ENSG00000260285 has higher accuracy when serving as a diagnosing marker of the liver cancer. According to the gene disclosed by the invention, in-vitro experiments find that restraining expression of the ENSG00000260285 can affect physiological characteristics of liver cancer cells, so that the gene can serve as a potential target to be applied to treating liver cancer.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to a gene related to liver cancer and its application, in particular to the ENSG00000260285 gene related to liver cancer and its application. Background technique [0002] Primary hepatocellular carcinoma (HCC), a human tumor of the digestive system, has a poor 5-year survival rate and is the second leading cause of cancer-related death worldwide. Hepatocellular carcinoma is highly morbid and fatal. It is a major global health challenge and the lack of early diagnostic tools is one of the clinical barriers to its effective treatment of HCC. [0003] Although there are new advances in surgical techniques and medical treatments, previous studies have identified many abnormally expressed protein-coding genes in HCC, and new molecular markers can play a role in early diagnosis, risk assessment is still urgently needed. Understanding the relationship between clinical symptoms and molecular chan...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68A61K45/00A61K49/00A61P35/00
CPCA61K45/00A61K49/0008C12Q1/6886C12Q2600/136C12Q2600/158C12Q2600/178
Inventor 董东石小峰
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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