Application of LINC01843 as liver cancer diagnosis and treatment marker

A technology for liver cancer and treatment of liver cancer, which is applied in the application field of liver cancer diagnosis and treatment markers, can solve problems such as functions that need to be further clarified, and achieve the effect of improving accuracy

Inactive Publication Date: 2020-08-11
TAIZHOU MUNICIPAL HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Compared with protein coding sequences and microRNA, lncRNA research is still in its infancy, and its function needs to be further elucidated

Method used

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  • Application of LINC01843 as liver cancer diagnosis and treatment marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Example 1 QPCR detection of differential expression of LINC01843

[0061] 1. RNA extraction

[0062] Tissue RNA was extracted using QIAGEN tissue RNA extraction kit, and the specific steps in the instructions were followed.

[0063] 2. Reverse transcription reaction

[0064] 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.

[0065]3. Real-time fluorescent quantitative PCR

[0066] PCR amplification was performed using cDNA as a template. The upstream primer of LINC01843 is 5′-GACTTGCTAATGGTGGAA-3′ (SEQ ID NO.2), the downstream primer is 5′-AAAGTGCTGGGATTATAGG-3′ (SEQ ID NO.3); GAPDH is an internal reference, and the upstream primer is 5′-CCGGGAAACTGTGGCGTGATGG-3′ (SEQ ID NO.4), the downstream primer is 5'-AGGTGGAG...

Embodiment 2

[0071] Example 2 Effect of LINC01843 gene on the proliferation of liver cancer cells

[0072] 1. Cell culture and cell transfection

[0073] Human hepatoma cell SMMC-7721 cells were inoculated in DMEM medium containing 10% fetal bovine serum in 5% CO 2 , Cultured in a constant temperature incubator at 37°C. 24h before transfection press 3×10 5 Inoculate each well in a 6-well plate, and add 2 mL of serum-free medium to make the cell density about 60% during transfection. Add the siRNA LipofectamineTM2000 complex (the configuration of the complex is operated according to the instructions) into a 6-well plate, the transfection system is 500 μL, and the final concentration of siRNA is 50 nmol / L. The experimental group is si-LINC01843 group, and the negative control group is nonsense Transfection of siRNA expression vector, ie si-NC group. The interference efficiency of siRNA was detected by QPCR, and the result showed that the interference efficiency of siRNA used in the prese...

Embodiment 3

[0080] Example 3 Soft agar colony formation experiment

[0081] 1. Experimental steps

[0082] (1) Digest the cells with 0.25% trypsin, pipette gently to make a single cell suspension, and collect the cell pellet by centrifugation.

[0083] (2) Resuspend with DMEM complete medium containing 20% ​​fetal bovine serum, count after appropriate dilution, and adjust the cell concentration to 5×10 3 pieces / ml.

[0084] (3) Prepare two low-melting-point agarose solutions with concentrations of 1.2% and 0.7%, respectively, and maintain them in a 40° C. water bath after autoclaving.

[0085] (4) Mix 1.2% agarose and 2×DMEM medium 1:1, add 2×antibiotics and 20% calf serum, take 3ml of the mixed solution and inject it into a 6cm-diameter plate, let it cool and solidify for 5 minutes, and place it as the bottom agar CO 2 Reserve in the incubator.

[0086] (5) Mix 0.7% agarose and 2×DMEM medium 1:1 in a sterile test tube, then add 0.2ml to the tube with a concentration of 5×10 3 cells...

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Abstract

The invention discloses an application of LINC01843 as a liver cancer diagnosis and treatment marker. According to the invention, up-regulation of expression of LINC01843 in liver cancer patients is discovered for the first time, and it is prompted that the LINC01843 can be used as a biomarker for diagnosis and treatment of liver cancer. Experiments further prove that through inhibition of expression of the LINC01843, proliferation, clone formation colony number and invasion and metastasis capabilities of liver cancer cells can be remarkably inhibited, and it is prompted that intervention of LINC01843 expression may become a new way for liver cancer treatment, and meanwhile, a theoretical basis is provided for research of a liver cancer mechanism.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to the application of LINC01843 as a marker for the diagnosis and treatment of liver cancer. Background technique [0002] Primary liver cancer is one of the most common malignant tumors in the world, its incidence ranks sixth among all cancers, and its mortality rate ranks second among all cancers. According to pathological classification, primary liver cancer can be divided into different types such as hepatocellular carcinoma (HCC), intrahepatic cholangiocarcinoma, and mixed hepatocellular carcinoma-intrahepatic cholangiocarcinoma. More than 90% of primary liver cancers are hepatocellular carcinomas. The occurrence of liver cancer is a multi-step and complicated process, mostly based on the chronic inflammation of the liver and the changed liver microenvironment, the mutation of many oncogenes or tumor suppressor genes, especially the gradual changes in their expression, and eventually ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/113A61K45/00A61K31/7105A61P35/00A61P1/16
CPCA61K31/7105A61K45/00A61P1/16A61P35/00C12Q1/6886C12Q2600/136C12Q2600/158C12Q2600/178
Inventor 王冬国冯继红
Owner TAIZHOU MUNICIPAL HOSPITAL
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