Application of linc00467 gene-targeted siRNA to leukemia medicine resistance

A technology of si-linc00467-r and si-linc00467-f, which is applied to medical preparations containing active ingredients, drug combinations, antineoplastic drugs, etc., can solve problems such as poor prognosis and survival rate, and achieve broad application prospects Effect

Active Publication Date: 2020-06-02
DONGGUAN EIGHTH PEOPLES HOSPITALDONGGUAN CHILDRENS HOSPITAL +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The current standard treatment plan is to induce remission with high-dose conventional chemotherapy, and follow-up treatment after complete remission (CR) is achieved to achieve the effect of intensive and maintenance treatment, but the prognosis and survival of this treatment rate is still not ideal

Method used

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  • Application of linc00467 gene-targeted siRNA to leukemia medicine resistance
  • Application of linc00467 gene-targeted siRNA to leukemia medicine resistance
  • Application of linc00467 gene-targeted siRNA to leukemia medicine resistance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1 Study on the expression level of long non-coding RNA linc00467 in leukemia patients

[0021] 1. Research object:

[0022] The research subjects included 16 patients with primary leukemia, 14 patients with normal leukemia, 11 patients with refractory leukemia and the leukemia cell line K562.

[0023] 2. Collection and storage of bone marrow mononuclear cells:

[0024] (1) Routinely extract 4ML of bone marrow fluid from each patient into a heparin anticoagulant tube.

[0025] (2) Take an empty tube, add 3ML of lymphocyte separation solution, slowly add an equal amount of bone marrow sample along the tube wall, and centrifuge at 1500rpm for 20 minutes. Pipette the mononuclear cells separated from the interface layer into a 1MLEP tube, add 1mL of 0.84% ​​ammonium chloride, mix well, let stand for 10 minutes to fully lyse residual red blood cells, and centrifuge at 1500rpm for 5 minutes.

[0026] (3) Discard the supernatant, add 1ML physiological saline, wash th...

Embodiment 2

[0034] Example 2 Correlation between the expression level of long-chain non-coding RNA linc00467 and the prognosis of leukemia patients

[0035] Check the tumor database TGCA for information related to the expression of linc00467 and prognosis of leukemia patients (such as figure 2 shown), it was found that the expression level of linc00467 was positively correlated with the prognosis of leukemia patients, and the survival time of patients with high expression of linc00467 was significantly shorter than that of patients with low expression of linc00467.

Embodiment 3

[0036] Example 3 Study on the interference efficiency of linc00467 using siRNA interference in leukemia cells K562

[0037] siRNA was transfected with a dose of 20nM to cultured K562 cells, and 48 hours later, total cellular RNA was extracted, reverse-transcribed into cDNA, and the expression level of linc00467 was detected by fluorescent quantitative PCR (experimental results are shown in image 3 shown), it can be seen that the siRNA has an interference efficiency of about 80% on linc00467. The siRNA sequence targeting the linc00467 gene is: si-linc00467-F: 5'-CAGGGAGGUUUAAUAGACATT-3', si-linc00467-R: 5'-UGUCUAUUAAACCUCCCUGTT-3'; the negative control siRNA sequence is: si-NC-F: 5 '-UUCUCCGAACGUGUCACGUTT-3', si-NC-R:5'-ACGUGACACGUUCGGAGAATT-3'.

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Abstract

The invention relates to the technical field of biological medicine, in particular to application of linc00467 gene-targeted siRNA to leukemia medicine resistance. According to the application, the siRNA is designed and synthesized by aiming at a nucleotide sequence of a linc00467 gene; and after the siRNA is transferred into a human leukemia cell K562, the expression of the linc00467 gene can beefficiently inhibited, and the medicine resistance of the K562 cell is obviously reduced, so that the anti-tumor purpose is achieved. Therefore, a linc00467 siRNA molecule designed by the invention can be used for preparing medicine for treating leukemia, and has wide application prospects.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to the application of siRNA targeting linc00467 gene in leukemia drug resistance. Background technique [0002] Leukemia is an aggressive hematological malignancy and the most common cause of cancer death among people aged 15 to 29, caused by the accumulation of immature leukemia cells in the blood and bone marrow. The current standard treatment plan is to induce remission with high-dose conventional chemotherapy, and follow-up treatment after complete remission (CR) is achieved to achieve the effect of intensive and maintenance treatment, but the prognosis and survival of this treatment rate is still not ideal. Leukemia drug resistance refers to the insensitivity or resistance of leukemia cells to the chemotherapy drugs used. When a patient undergoes several courses of combined chemotherapy, the percentage of leukemia cells in the bone marrow does not decrease significantly, ...

Claims

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Application Information

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IPC IPC(8): A61K31/713A61P35/02C12Q1/6886
CPCA61K31/713A61P35/02C12Q1/6886C12Q2600/118C12Q2600/166C12Q2600/158C12Q2600/178
Inventor 饶春宝陆小梅
Owner DONGGUAN EIGHTH PEOPLES HOSPITALDONGGUAN CHILDRENS HOSPITAL
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