Use of lncRNA as biomarker in diagnosis and treatment of liver cancer

A technology for liver cancer and liver cancer treatment, applied in the field of biomedicine, can solve the problem that the function needs to be further clarified, and achieve the effect of improving the accuracy rate

Active Publication Date: 2017-10-20
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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  • Use of lncRNA as biomarker in diagnosis and treatment of liver cancer
  • Use of lncRNA as biomarker in diagnosis and treatment of liver cancer
  • Use of lncRNA as biomarker in diagnosis and treatment of liver cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1 Screening Gene Markers Related to Liver Cancer

[0059] 1. Sample collection

[0060] 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.

[0061] 2. Preparation of RNA samples

[0062] Tissue RNA was extracted using QIAGEN tissue RNA extraction kit, and the operation was performed according to the specific steps in the manual.

[0063] 3. Reverse transcription and labeling

[0064] 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.

[0065] 4. Hybridization

[0066] The gene chip uses Cannes Bio-Human lncRNA Array, and hybridization is performed according to the steps of the chip instruction manual.

[0067] 5. Data processing

[0068] After hybridization,...

Embodiment 2

[0071] Example 2 QPCR sequencing verification of differential expression of LOC101927480 gene

[0072] 1. Large-scale QPCR verification of the differential expression of the LOC101927480 gene. According to the sample collection method in Example 1, 60 samples of liver cancer tissues and 60 samples of paracancerous tissues were collected.

[0073] 2. The RNA extraction steps are the same as in Example 1.

[0074] 3. Reverse transcription:

[0075] 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.

[0076] (3) QPCR amplification test

[0077] Primer design:

[0078] The primer sequence of LOC101927480 gene is:

[0079] Forward primer: 5'-AATCTAGGACTTACGCTCTT-3' (SEQ ID NO.2)

[0080] Reverse primer: 5'-CACTGAATGG...

Embodiment 3

[0090] Example 3 Differential expression of LOC101927480 gene in liver cancer cell lines

[0091] 1. Cell culture

[0092] 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.

[0093] 2. Extraction of RNA

[0094] 1) Stop the culture when the cells reach 80-90% confluency, digest with 0.25% trypsin and collect the cells in 1.5ml EP tubes, add lm1Trizol to each tube and shake slowly to break the cells, place on ice for 10min.

[0095] 2) Remove protein and DNA: add 0.2ml chloroform to each 1.5ml EP tube, shake for 15s, and place at room temperature for 10min. Centrifuge at 12000rpm for 15min at 4°C.

[0096] The rest of the operation steps are the same as the RNA extraction process from the ...

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Abstract

The invention discloses a use of lncRNA as a biomarker in the diagnosis and treatment of liver cancer. The marker is LOC101927480 concretely. The up-regulation of the expression of the LOC101927480 in liver cancer patients is found for the first time, so the LOC101927480 can be used as a biomarker in the diagnosis and treatment of the liver cancer. Experiments further confirm that the inhibition of the LOC101927480 expression can significantly reduce the proliferation and clone formed colony number of liver cancer cells, so the intervention of the LOC101927480 expression may become a new way for the treatment of the liver cancer. The use provides a theoretical basis for the study of the liver cancer mechanism.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to the use of lncRNA as a biomarker in the diagnosis and treatment of liver cancer, in particular to the biomarker LOC101927480. 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 ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68A61K31/7105A61K48/00A61P35/00
CPCA61K31/7105C12Q1/6886C12Q2600/158C12Q2600/178
Inventor 王冬国陈佳玉杨林军
Owner TAIZHOU MUNICIPAL HOSPITAL
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