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Applications of LINC01426 in hepatocellular carcinoma diagnosis and therapy

A technology of 1.LINC01426, a.linc01426, applied in medical preparations containing active ingredients, recombinant DNA technology, biochemical equipment and methods, etc., can solve the large heterogeneity of liver cancer, poor prognosis, poor patient treatment effect, etc. problem, to achieve the effect of improving the accuracy

Active Publication Date: 2018-06-15
TAIZHOU MUNICIPAL HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although with the continuous development and progress of medical technology, the comprehensive treatment of liver cancer has made great achievements, but because most patients are already in the middle and late stages when they are diagnosed and the heterogeneity of liver cancer is large, the treatment effect of most patients is poor and the prognosis is poor.

Method used

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  • Applications of LINC01426 in hepatocellular carcinoma diagnosis and therapy
  • Applications of LINC01426 in hepatocellular carcinoma diagnosis and therapy
  • Applications of LINC01426 in hepatocellular carcinoma diagnosis and therapy

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] Example 1 Screening Gene Markers Related to Liver Cancer

[0052] 1. Sample collection

[0053] The cancer tissues and paracancerous tissues of 5 liver cancer patients were collected, and the patients gave informed consent, and all the above specimens were obtained with the consent of the organizational ethics committee.

[0054] 2. Preparation of RNA samples

[0055] Tissue RNA was extracted using QIAGEN tissue RNA extraction kit, and the operation was performed according to the specific steps in the manual.

[0056] 3. Quality analysis of RNA samples

[0057] Nanodrop2000 was used to detect the concentration and purity of the extracted RNA, agarose gel electrophoresis was used to detect RNA integrity, and Agilent2100 was used to determine the RIN value. Concentration ≥ 200ng / μl, OD260 / 280 between 1.8 and 2.2.

[0058] 4. Remove rRNA

[0059] Ribosomal RNA was removed from total RNA using the Ribo-Zero kit.

[0060] 5. Construction of cDNA library

[0061] The c...

Embodiment 2

[0068] Example 2 QPCR sequencing to verify the differential expression of the LINC01426 gene

[0069] 1. Large-scale QPCR verification of the differential expression of the LINC01426 gene. According to the sample collection method in Example 1, 60 samples of liver cancer tissues and 60 samples of paracancerous tissues were collected.

[0070] 2. RNA extraction The steps are the same as in Example 1.

[0071] 3. Reverse transcription:

[0072] 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.

[0073] (3) QPCR amplification test

[0074] Primers were designed according to the sequences of LINC01426 gene and housekeeping gene GAPDH, and the primer sequences were synthesized by Shanghai Sangong. The designed primer ...

Embodiment 3

[0087] Example 3 Differential expression of LINC01426 gene in liver cancer cell lines

[0088] 1. Cell culture

[0089] Human liver cancer cell lines HepG2, Huh7 and normal liver cell line HL-7702 were incubated in DMEM containing 10% fetal bovine serum and 1% P / S at 37°C and 5% CO 2 , Cultivated in an incubator with a relative humidity of 90%. Change the medium once every 2-3 days, and use 0.25% EDTA-containing trypsin for routine digestion and passage.

[0090] 2. Extraction of RNA

[0091] The RNA in the cells was extracted using Qiagen's cellular RNA extraction kit, and the experimental operation was performed according to the instructions.

[0092] 3. The specific steps of reverse transcription are the same as in Example 2

[0093] 4. Statistical methods

[0094] The experiments were all repeated 3 times, and the result data were expressed in the form of mean ± standard deviation. SPSS18.0 statistical software was used for statistical analysis, and the difference betwe...

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Abstract

The invention discloses a biological marker used for liver cancer diagnosis and treatment, and more specifically discloses marker LINC01426. It is found for the first time that differential expressionof LINC01426 is related to generation and development of liver cancer; in patients with liver cancer, expression up-regulation of LINC01426 is detected, it reveal that intervention on LINC01426 geneexpression could be a novel liver cancer treatment method. It is found in verification tests that reduction of LINC01426 expression with inhibitors is capable of reducing propagation and cell formation clone colony number of liver cancer cells obviously.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to the application of LINC01426 in the diagnosis and treatment of hepatocellular carcinoma. Background technique [0002] Liver cancer is a major disease that seriously threatens the lives and health of our people. Its morbidity and mortality have long been ranked fourth and second among malignant tumors in my country. Primary liver cancer is mainly divided into three subtypes: hepatocellular carcinoma, which is the most important subtype, accounts for about 85% of primary liver cancer; cholangiocellular carcinoma, as the second largest type, has the highest incidence rate. increasing; and mixed liver cancer, including hepatocellular carcinoma and cholangiocarcinoma. Although with the continuous development and progress of medical technology, the comprehensive treatment of liver cancer has made great achievements, but because most patients are already in the middle and late stages when the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886A61K31/7105A61P35/00C12N15/113
CPCA61K31/7105C12Q1/6886C12Q2600/178
Inventor 牟永华王冬国刑人伟
Owner TAIZHOU MUNICIPAL HOSPITAL
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