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Long noncoding RNA (lncRNA) marker associated with hepatocellular carcinoma, and detection primer and application thereof

A technology of hepatocellular carcinoma and primer sequence, which is applied in the field of biomedicine and can solve the problems such as the lack of unified molecular marker standards for hepatocellular carcinoma

Active Publication Date: 2019-04-12
TAISHAN MEDICAL UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is no uniform molecular marker standard for HCC

Method used

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  • Long noncoding RNA (lncRNA) marker associated with hepatocellular carcinoma, and detection primer and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 Screening of gene markers related to hepatocellular carcinoma

[0044] 1. Sample collection

[0045] The cancer tissues and corresponding para-cancerous tissue samples from 27 patients with primary hepatocellular carcinoma were collected, and 5 samples were randomly selected for high-throughput sequencing. The patients had informed consent. All the above-mentioned samples were obtained by the organization ethics committee. agree.

[0046] 2. Preparation and quality analysis of RNA samples

[0047] Use TRIZOL method to extract total RNA from tissue

[0048] 1) Cut the tissue into pieces with scissors, add 1ml Trizol, shake on a shaker for 1 min; place it at room temperature for 10 min to completely decompose the nucleoprotein body.

[0049] 2) Add 200μl of chloroform (chloroform), close the tube cap, shake vigorously for 15s, and let it stand for 10min at room temperature.

[0050] 3) Centrifuge at 11000rpm for 15min at 4°C.

[0051] 4) Transfer the water sample layer to a...

Embodiment 2

[0078] Example 2 QPCR sequencing to verify the differential expression of RP11-284F21.10 gene

[0079] 1. The differential expression of RP11-284F21.10 gene was verified by large sample QPCR using the cancer tissue samples and adjacent tissue samples collected from 27 patients.

[0080] 2. RNA extraction

[0081] Using Trizol method to extract tissue RNA, see Example 1 for specific steps.

[0082] 3. QPCR detection

[0083] 1) Primer design

[0084] The primers are designed according to the gene sequence of RP11-284F21.10 and GADPH. The specific primer sequence is as follows:

[0085] RP11-284F21.10 gene:

[0086] The forward primer is 5'-GAGCCTACACAATATAAGC-3' (SEQ ID NO. 2);

[0087] The reverse primer is 5'-TAACAGCATCATCCACAT-3' (SEQ ID NO. 3).

[0088] GAPDH gene:

[0089] The forward primer is 5'-AATCCCATCACCATCTTCCAG-3' (SEQ ID NO. 4);

[0090] The reverse primer is 5'-GAGCCCCAGCCTTCTCCAT-3' (SEQ ID NO. 5).

[0091] 2) Reverse transcription reaction

[0092] Use FastQμant cDNA First Strand...

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Abstract

The invention discloses a long noncoding RNA (lncRNA) marker associated with hepatocellular carcinoma as well as a detection primer and application thereof. The lncRNA marker associated with hepatocellular carcinoma is RP11-284F21.10. By performing high throughput sequencing, significant up-regulation of RP11-284F21.10 expression in patients with hepatocellular carcinoma is discovered for the first time by the invention; moreover, up-regulation of RP11-284F21.10 expression in patients with hepatocellular carcinoma is further verified by performing QPCR, suggesting that the RP11-284F21.10 can be used as a biomarker in diagnosis and treatment of hepatocellular carcinoma.

Description

Technical field [0001] The present invention belongs to the field of biomedicine, and relates to lncRNA markers related to liver cancer and detection primers and applications thereof. The marker is RP11-284F21.10. Background technique [0002] Liver cancer is a common clinical malignant tumor, which is widely distributed and spread all over the world. According to the cause of liver cell canceration, liver cancer is generally divided into primary liver cancer and secondary liver cancer. Secondary liver cancer mainly refers to the tumor tissue in the liver caused by the transfer of body fluids or infiltration of the liver after other organs become cancerous, and it is more common in cancer patients such as stomach cancer and bowel cancer. Compared with secondary liver cancer, primary liver cancer refers to cancers whose cell carcinogenesis originates in the liver, and is the main research object for studying the pathogenesis and development of liver cancer. Primary liver cancer ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12N15/113
CPCC12Q1/6886C12Q2600/178C12Q2600/158
Inventor 李正美赵强邱建峰侯坤石丽婷赵慧慧路伟钊
Owner TAISHAN MEDICAL UNIV
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