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Long non-coding RNA LINC02481 related to gastric cancer and application of long non-coding RNA LINC02481

A LINC02481, long-chain non-coding technology

Active Publication Date: 2020-06-16
PEKING UNIV SHENZHEN HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Many patients with early gastric cancer lack specific clinical symptoms, and patients usually do not pay attention to them, which leads to the neglect of gastric cancer

Method used

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  • Long non-coding RNA LINC02481 related to gastric cancer and application of long non-coding RNA LINC02481
  • Long non-coding RNA LINC02481 related to gastric cancer and application of long non-coding RNA LINC02481
  • Long non-coding RNA LINC02481 related to gastric cancer and application of long non-coding RNA LINC02481

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Detection of the expression difference of LINC02481 in gastric cancer tissues and adjacent tissues

[0027] 1. Research object

[0028] 20 cases of gastric cancer tissues and 20 cases of corresponding paracancerous tissues were obtained from Qingdao Cancer Hospital. All tissue samples were reviewed and approved by the Hospital Ethics Committee, and all tissue samples were obtained from patients who signed informed consent. The gastric cancer tissue was confirmed to be gastric cancer by pathological examination (when the gastric cancer tissue was resected, the patient had not yet received chemotherapy, radiotherapy and other treatments).

[0029] The specific organization is as follows

[0030]

[0031]

[0032] 2. Tissue RNA Extraction

[0033] (1) Put 50 mg of gastric cancer tissue or paracancerous tissue into a mortar, add a small amount of liquid nitrogen, and grind rapidly. After the tissue becomes soft, add a small amount of liquid nitrogen for grinding aga...

Embodiment 2

[0068] Real-time quantitative PCR detection of LINC02481 gene expression after knockdown

[0069] (1) Cell culture

[0070] Human gastric cancer cell MKN-45 was cultured in high-glucose DMEM medium (10% fetal bovine serum) at 37°C, 5% CO 2 Constant temperature cell incubator culture.

[0071] (2) si-RNA transfection

[0072] (1) The day before transfection, 2×10 5 MKN-45 cells were seeded in 6-well plates at 37 °C, 5% CO 2 Culture in a constant temperature cell incubator for 24 hours, and transfect according to the instructions of Lipofectamine2000;

[0073] (2) The experimental groups were control group, si-NC group, si-LINC02481-1, si-LINC02481siRNA-2, si-LINC02481-3.

[0074] 5'-AUUUGUCGGAGGCAUUAUCAA-3' (SEQ ID NO: 6)

[0075] 5'-GAUAAUGCCUCCGACAAAUAU-3' (SEQ ID NO: 7)

[0076] si-LINC02481-2

[0077] 5'-GAAGGUGUUUGCUAGACAAAG-3' (SEQ ID NO:8)

[0078] 5'-UUGUCUAGCAAACACCUUCUG-3' (SEQ ID NO:9)

[0079] si-LINC02481-3

[0080] 5'-UCAUGAUAAAUCGAACAAGCA-3' (SEQ ID NO...

Embodiment 3

[0089] CCK-8 assay detects the effect of transfection si-LINC02481-2 on the proliferation ability of gastric cancer cell MKN-45

[0090] experimental method

[0091] (1) will 2×10 3 The MKN-45 cells were seeded in 96-well plates, 100 μl per well, and three replicate wells were set up in each group, and cultured in a 5% CO2 incubator at 37°C;

[0092] (2) MKN-45 cells were transfected with si-NC and si-LINC02481-2 respectively. 4 hours before the end of the culture at 24h, 48h, and 72h, 10ul CCK-8 was added to each well, and after 4h incubation in a constant temperature incubator, 450nm Detect absorbance.

[0093] Experimental results

[0094] pass Figure 4 It can be seen that at 24 hours of transfection, there was no significant difference between the si-NC group and the si-LINC02481-2 group, but at 48 and 72 hours (48 hours: si-NC 0.649±0.035, si-LINC02481-2 0.431±0.027; 72h: si-NC 1.057±0.415, si-LINC02481-2 0.683±0.039), the OD value of MKN-45 cells transfected with s...

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Abstract

The invention aims to provide a long-chain non-coding RNA LINC02481 related to gastric cancer and an application of the long-chain non-coding RNA LINC02481. The long-chain non-coding RNA provided by the invention is highly expressed in gastric cancer tissues, so that the long-chain non-coding RNA can be used as a target for treating gastric cancer. The siRNA inhibitor designed according to the LINC02481 can inhibit the proliferation of gastric cancer cells and promote the apoptosis of the gastric cancer cells, so that the siRNA provided by the invention can be applied to the preparation of gastric cancer drugs. The invention has the beneficial effects that the invention provides a new target spot for gastric cancer diagnosis, and the detection has excellent diagnostic value; secondly, theinvention provides the application of the gene inhibitor of LINC02481 in preparation of the medicine for treating the gastric cancer, and a new direction is provided for development of the medicine for treating the gastric cancer.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a long non-coding RNA LINC00248 related to gastric cancer and its application. Background technique [0002] Gastric cancer is a common malignant tumor of the digestive system. Because the pathogenesis of gastric cancer is very complicated, and the early symptoms of patients are not obvious, the early diagnosis rate of gastric cancer patients is low. Gastric cancer is the fourth largest malignant tumor in the world. Although the incidence of gastric cancer has decreased in recent years, the mortality rate of patients has not been effectively reduced. In China, the mortality rate of gastric cancer patients still ranks second among all tumors. Gastric cancer is divided into early gastric cancer and advanced gastric cancer. Most gastric cancer patients are already in the advanced stage when they are diagnosed. Many patients with early gastric cancer lack specific cl...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886A61K45/00A61K31/7088A61P35/00A61P1/00
CPCA61K31/7088A61K45/00A61P35/00C12Q1/6886C12Q2600/158C12Q2600/178
Inventor 王颖翠
Owner PEKING UNIV SHENZHEN HOSPITAL
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