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Long non-coding RNA (lncRNA) and application thereof to diagnosis/treatment of gastric cancers

A non-coding, gastric cancer technology applied in the field of genetic engineering

Inactive Publication Date: 2016-11-09
JIANGSU PROVINCE HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the functions and mechanisms of some lncRNAs have been reported in gastric cancer, the role of other lncRNAs in the development of gastric cancer still needs further study

Method used

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  • Long non-coding RNA (lncRNA) and application thereof to diagnosis/treatment of gastric cancers
  • Long non-coding RNA (lncRNA) and application thereof to diagnosis/treatment of gastric cancers
  • Long non-coding RNA (lncRNA) and application thereof to diagnosis/treatment of gastric cancers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0220] Example 1 Detecting the expression of lncRNA DUXAP8 in tissues and cells

[0221] Take 0.1g tissue, grind it sufficiently with liquid nitrogen (into powder) or 1-5×10 7 Discard the culture medium from the cells, and rinse twice with pre-cooled PBS. Add 1ml of Trizol lysate, pipette and mix with an enzyme-free pipette tip, let stand for 5min, and transfer the lysate into a pre-labeled 1.5ml enzyme-free centrifuge tube. Centrifuge at 7500g at 4°C for 5 minutes, take the supernatant and add 1 / 5 volume of chloroform, invert and mix for 30 seconds, and let stand for 2 minutes. Centrifuge at 12000g for 15min at 4°C. The solution is divided into three layers (aqueous phase-white precipitate-red organic matter), transfer the aqueous phase layer to a new 1.5ml centrifuge tube, try not to absorb the white precipitate. Add an equal volume of isopropanol, mix gently by inversion, and let stand for 5-10min. Centrifuge at 12000g for 10min at 4°C. Discard the supernatant, add 1ml...

Embodiment 2

[0240] Example 2 Highly expressed lncRNA DUXAP8 is significantly correlated with gastric cancer tumor size, lymph node metastasis, clinical classification and poor prognosis

[0241] In order to detect the correlation between lncRNA DUXAP8 expression and clinicopathological features, gastric cancer patients were divided into two groups according to the median value of relative lncRNA DUXAP8 expression in tumor tissue: a relatively low expression group of lncRNA DUXAP8 and a relatively high expression group of lncRNA DUXAP8 ( figure 1 C). Notably, high expression of lncRNA DUXAP8 in gastric cancer was significantly correlated with TNM stage (p=0.001), lymph node metastasis (P=0.007) and tumor size (P=0.002). However, the expression of lncRNA DUXAP8 was not correlated with other parameters such as gender (P=0.635) and age (P=0.157). To determine the relationship between the expression of lncRNA DUXAP8 and the prognosis of gastric cancer patients, progression-free survival (PFS)...

Embodiment 3

[0242] Example 3 Effect of lncRNA DUXAP8 on the proliferation of gastric cancer cells

[0243] MTT experiment

[0244] SGC-7901 and BGC-823 cells 24 hours after transfection were inoculated in 96-well culture plates at 3000 cells per well; after 80% of the cells adhered to the wall, the cells were synchronized for 12 hours, and the original medium was discarded. Six replicate wells were set up for each sample, and the total reaction volume in each well was 200 μl. Add 20 μl of MTT reaction solution (5 mg / ml, dissolved in PBS) to each well, incubate at 37°C in the dark for 4 hours; discard the supernatant, add 150 μl dimethyl sulfoxide (DMSO) to each well, shake on a micro-shaker After 10 min, the absorbance at a wavelength of 490 nm was measured with a microplate reader.

[0245] Clonogenic experiment

[0246] Digested with 0.25% trypsin and blown into a single cell suspension, inoculated on a 6-well plate with an appropriate cell density (500 cells) to disperse the cells e...

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Abstract

The invention belongs to the field of genetic engineering and in particular relates to an application of long non-coding RNA (lncRNA) DUXAP8 to prognosis of gastric cancers and target drug treatment. With up-regulation of IncRNA DUXAP8 in gastric cancer cells, prognosis of patients with IncRNA DUXAP8 with high expression level is relatively poor. Effects are exerted on invasion, migration, and the like of the gastric cancer cells by changing expression of IncRNA DUXAP8. Gastric cancer cell proliferation and apoptosis induction can be inhibited by knocking down IncRNA DUXAP8 expression.

Description

technical field [0001] The invention belongs to the field of genetic engineering, in particular to a long non-coding RNA and its application in diagnosis / treatment of gastric cancer. Background technique [0002] Gastric cancer is one of the most common malignant gastrointestinal tumors in China, and nearly half of gastric cancer patients in the world are in China. Although the development of comprehensive treatment methods such as surgery, intervention, radiotherapy and chemotherapy, and targeted therapy has led to a downward trend in morbidity and mortality, it is still the second leading cause of cancer death. Early gastric cancer can be cured by endoscopic mucosal resection and radical gastrectomy, but the vast majority of gastric cancer patients are already in the middle and late stage of the disease when they are first diagnosed. High risk of recurrence and metastasis, the prognosis is still not optimistic. The high morbidity and mortality of gastric cancer not only ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12Q1/68A61K48/00A61K31/7088A61P35/00
CPCA61K31/7088C12N15/113C12N2310/10C12Q1/6886C12Q2600/118C12Q2600/136C12Q2600/158C12Q2600/178
Inventor 张智弘马红伟
Owner JIANGSU PROVINCE HOSPITAL
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