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Long non-coding RNA (Ribose Nucleic Acid) and application thereof to diagnosis/treatment of non-small-cell lung cancer

A non-small cell lung cancer, non-coding technology, used in DNA/RNA fragments, recombinant DNA technology, medical preparations containing active ingredients, etc., can solve problems such as poor prognosis

Inactive Publication Date: 2016-09-21
JIANGYIN PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

We further found that downregulation of LINC00961 in NSCLC was associated with larger tumor volume and higher clinical stage; patients with low expression level of LINC00961 had relatively poor prognosis

Method used

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  • Long non-coding RNA (Ribose Nucleic Acid) and application thereof to diagnosis/treatment of non-small-cell lung cancer
  • Long non-coding RNA (Ribose Nucleic Acid) and application thereof to diagnosis/treatment of non-small-cell lung cancer
  • Long non-coding RNA (Ribose Nucleic Acid) and application thereof to diagnosis/treatment of non-small-cell lung cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] Example 1 Detection of the expression of LINC00961 in tissues and cells

[0072] Take 0.1g tissue, grind it sufficiently with liquid nitrogen (into powder) or 1-5×10 7Discard 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 o...

Embodiment 2

[0093] Example 2 Effect of LINC00961 on proliferation of NSCLC cells

[0094] In order to study the functional role of LINC00961 in NSCLC cells, first, we used qRT-PCR to detect the expression of LINC00961 in different NSCLC cell lines. The specific method is the same as before. Such as figure 2 As shown in A, LINC00961 expression was significantly downregulated in A549 and PC9 cells when compared to a normal bronchial epithelial cell line (16HBE). To control LINC00961 levels in NSCLC cells, pcDNA-LINC00961 vector and empty vector were transfected into A549 and PC9 cells. After 48 hours of transfection, QRT-PCR analysis showed that the expression of LINC00961 was up-regulated in cells transfected with pcDNA-LINC00961 vector ( figure 2 B). Next, the MTT assay was used to detect the effect of overexpressing LINC00961 on the growth of A549 and PC9 cells. The specific steps are as follows:

[0095] 1) Cell inoculation: Digest monolayer cultured cells with 0.25% trypsin, and...

Embodiment 3

[0101] Example 3 Overexpression of LINC00961 inhibits NSCLC cell invasion and migration

[0102] In addition to cell proliferation, cell invasion and migration also play an important role in the development and progression of tumors. Therefore, we used Transwell chamber experiments to evaluate the effect of LINC00961 on the invasion and migration of NSCLC cells. Specific steps are as follows:

[0103] 1) Place a Transwell chamber with a pore size of 8 μm in a 24-well plate. For the cell invasion experiment, 50mg / l BD Matrigel 1:8 dilution was used to coat the upper chamber surface of the bottom membrane of the Transwell chamber, put the coated chamber into a 24-well plate, and incubate in the incubator for 2 hours.

[0104] 2) Digest the cells, centrifuge to discard the culture medium after the digestion is terminated, wash with PBS 1-2 times, and resuspend with BSA-containing serum-free medium. Adjust cell density to 1x10 5 . Take 200 μl of the cell suspension and add it...

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Abstract

The invention belongs to the field of gene engineering, and particularly relates to application of LINC00961 to prediction of non-small-cell lung cancer (NSCLC) prognosis and treatment of target spot medicine. In the NSCLC, down-regulation of the LINC00961 is closely related to clinical stages and tumor sizes, and is closely related to patient prognosis at the same time. The invasion, migration and the like of NSCLC cells are influenced by changing expression of the LINC00961, so that the LINC00961 expression is enhanced, and the invasion and migration of the NSCLC cells can be restrained.

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 non-small cell lung cancer. Background technique [0002] Lung cancer is the most common cause of tumor-related death worldwide, and non-small cell lung cancer (NSCLC) accounts for about 80%-85% of all lung cancer cases. Lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC) are the two main forms of NSCLC. Despite recent advances in the clinical treatment of NSCLC, the survival rate of patients with NSCLC remains disappointing. Therefore, a better understanding of the pathogenesis and molecular mechanisms of NSCLC is crucial for the diagnosis and treatment of NSCLC. [0003] Recently, a large number of proto-oncogenes and tumor suppressor genes have been identified as key regulators of NSCLC tumorigenesis and progression. In addition to protein-coding genes, lncRNA has also become a key molecule...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12Q1/68G01N33/574A61K31/7088A61P35/00
CPCA61K31/7088C12Q1/6886C12Q2600/118C12Q2600/136C12Q2600/158C12Q2600/178G01N33/57423
Inventor 刘志利茅卫东王赛花姜彬
Owner JIANGYIN PEOPLES HOSPITAL
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