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Long-chain non-coding RNA gene marker for detecting liver cancer and application of long-chain non-coding RNA gene marker for detecting liver cancer

A long-chain non-coding, liver cancer technology, applied in the field of detecting long-chain non-coding RNA gene markers of liver cancer, can solve problems such as lack of function, RNA formation protein, etc., and achieve the effect of improving survival rate and excellent diagnostic value.

Active Publication Date: 2020-05-22
宜兴市拜奥精核生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Only a small number of genes in the human genome can encode proteins, and 98% of the genes can be transcribed to form RNA, but these RNAs cannot further form proteins. The RNA formed by the transcription of these genes is called non-coding RNA. Before, people Treating these noncoding RNAs as transcriptional noise with no actual specific function

Method used

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  • Long-chain non-coding RNA gene marker for detecting liver cancer and application of long-chain non-coding RNA gene marker for detecting liver cancer
  • Long-chain non-coding RNA gene marker for detecting liver cancer and application of long-chain non-coding RNA gene marker for detecting liver cancer
  • Long-chain non-coding RNA gene marker for detecting liver cancer and application of long-chain non-coding RNA gene marker for detecting liver cancer

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Detection of the expression of LINC01141 in liver cancer tissues and adjacent tissues

[0029] 1. Research object

[0030] A total of 20 liver cancer tissues and corresponding paracancerous tissues were selected for the experiment. The tissues were obtained from Qingdao Cancer Hospital. All tissue samples were reviewed and approved by the Hospital Ethics Committee, and all patients who obtained the tissue samples signed informed consent. The liver cancer tissue was confirmed to be liver cancer by pathological examination (when the liver cancer tissue was resected, the patient had not yet received chemotherapy, radiotherapy and other treatments).

[0031] The specific specimens are as follows:

[0032]

[0033]

[0034]

[0035] 2. Tissue RNA Extraction

[0036] (1) Take out the tissue sample from the -80°C refrigerator, take about 50 mg of the tissue sample and put it into a mortar, add a small amount of liquid nitrogen, and grind it quickly. After the tissue...

Embodiment 2

[0067] Knockdown of the LINC01141 gene

[0068] 1. siRNA design

[0069] si-RNA-1

[0070] 5'-AUUUUGCAGAUCUAUCAUCCU-3' (SEQ ID NO.6)

[0071] 5'-GAUGAUAGAUCUGCAAAAUUC-3' (SEQ ID NO.7)

[0072] si-RNA-2

[0073] 5'-AUCAAGUAUCUAUAUGUGCCA-3' (SEQ ID NO.8)

[0074] 5'-GCACAUAUAGAUACUUGAUAC-3' (SEQ ID NO.9)

[0075] si-RNA-1 and si-RNA-2 were synthesized by Shanghai Gemma Company, and si-NC was provided by the company.

[0076] 2. Cell Culture

[0077] Human liver cancer cells HepG2 were cultured in high-glucose DMEM medium (10% fetal bovine serum), culture conditions: cell constant temperature incubator at 37°C, 5% CO 2 .

[0078] 3. siRNA transfection

[0079] (1) The day before transfection, 2×10 5 Seed HepG2 cells onto 6-well cell culture plates at 37 °C, 5% CO 2 After culturing in the incubator for 24 hours, the cells were transfected according to the instructions of Lipofectamine 2000;

[0080] (2) Experimental groups were divided into control group (empty transfe...

Embodiment 3

[0090] 1. Cell Proliferation Experiment

[0091] (1) Divide 5×10 3 HepG2 cells transfected with si-NC and si-RNA-1 were inoculated in a 96-well plate, 90 μl per well, with 3 replicate wells for each group, and placed at 37°C, 5% CO 2 cultured in a cell culture incubator;

[0092] (2) Detection was performed at 24h, 48h, and 72h respectively. During detection, 10ul of CCK-8 detection solution was added to each well, and then placed in a 37°C, 5% CO2 incubator for 4 hours;

[0093] (3) Place the cell culture plate in a microplate reader, detect the absorbance at 450 nm, and draw the growth curve.

[0094] 2. Results

[0095] pass Figure 4 It can be seen that after the experimental group was transfected with si-RNA-1, compared with the si-NC group, the proliferation of HepG2 cells was inhibited, and the proliferation rate was significantly reduced. At 72h, the OD value of the si-NC group was 1.09± 0.049, and the OD value of the si-RNA-1 group was 0.766±0.041, the difference...

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Abstract

The invention relates to a long-chain non-coding RNA gene marker for detecting liver cancer and an application of the long-chain non-coding RNA gene marker for detecting liver cancer. The long-chain non-coding RNA is LINC01141. Through fluorescence quantitative PCR detection, the inventor of the invention finds that the expression of the LINC01141 in liver cancer patients is notably raised, whichillustrates that the LINC01141 can be used as a gene marker of the liver cancer and can be used for diagnosing and treating the liver cancer. The long-chain non-coding RNA gene marker for detecting liver cancer disclosed by the invention has the beneficial effects that early diagnosis of the liver cancer can be realized through detecting the expression level of the LINC01141 in the liver cancer patients, so that the survival rate of the liver cancer patients can be increased. Secondly, the invention provides an application of LINC01141 siRNA to preparation of a medical composition for liver cancer. The medical composition can be used as a new treatment medicine for treating the liver cancer patients, and has important clinical value and application prospects.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a long-chain non-coding RNA gene marker for detecting liver cancer and its application. Background technique [0002] Liver cancer is one of the most common cancers in clinical practice. The annual death rate of liver cancer ranks third among malignant tumors in the world, and the new incidence rate ranks fifth among malignant tumors in the world, and the occurrence of liver cancer shows a trend of rapid increase. The pathogenesis of liver cancer is related to viral infection, non-alcoholic fatty liver, alcoholic hepatitis, genetic and environmental factors, etc. Most liver cancer patients are diagnosed and treated at advanced stages, so the survival rate of patients is low. Early diagnosis and early treatment are the key to improve the survival rate of liver cancer patients. [0003] Only a small number of genes in the human genome can encode proteins, and 98%...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12Q1/6886A61K31/713A61P1/16A61P35/00
CPCA61K31/713A61P1/16A61P35/00C12N15/113C12Q1/6886C12Q2600/158C12Q2600/178
Inventor 曹爱华
Owner 宜兴市拜奥精核生物科技有限公司
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