Application of long-chain non-coding RNA LAMP5-AS1 to MLL-R leukemia

A RNALAMP5-AS1, long-chain non-coding technology, applied in the field of biomedicine, can solve the problems that the degradation of lncRNA and MLL fusion protein has not been reported, and achieve the effect of prolonging the life cycle

Active Publication Date: 2020-10-02
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are few reports on lncRNA in MLL-R leukemia, and the

Method used

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  • Application of long-chain non-coding RNA LAMP5-AS1 to MLL-R leukemia
  • Application of long-chain non-coding RNA LAMP5-AS1 to MLL-R leukemia
  • Application of long-chain non-coding RNA LAMP5-AS1 to MLL-R leukemia

Examples

Experimental program
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Effect test

Embodiment 1

[0038] Example 1 LAMP5-AS1 expression analysis and evaluation of its clinical value

[0039] The present invention discovers an lncRNA LAMP5-AS1 that is specifically highly expressed in MLL-R leukemia and lowly expressed in normal physiological conditions and other types of leukemia. In order to further confirm the expression specificity of LAMP5-AS1 in MLL-R leukemia, we re-collected a batch of bone marrow samples from the First Affiliated Hospital of Sun Yat-sen University for detection and analysis, including MLL wild-type (MLL-wt) leukemia samples163 cases, and 53 cases of MLL-R leukemia samples. All sample collection was approved by the Ethics Committee of Sun Yat-Sen University and informed consent was obtained from the patients. LAMP5-AS1 was specifically detected by extracting RNA and using qRT-PCR technology. The primer sequences of the qRT-PCR used are as follows:

[0040] Forward primer sequence: 5'-CACTGAACGGATCTCAAACC-3' (SEQ ID NO: 2);

[0041] Reverse primer...

Embodiment 2

[0043] Example 2 Functional identification of LAMP5-AS1 in MLL-R leukemia

[0044] In order to solve the core problem of LAMP5-AS1 participating in the regulation of MLL-R leukemia, this example intends to further study the function of LAMP5-AS1 on MLL-R leukemia. MLL-R leukemia cell lines, THP1 (MLL-AF9) and MV4-11 (MLL-AF4) were selected as the research objects. Through siRNA interference technology, two different siRNA sequences were designed for LAMP5-AS1.

[0045] Forward sequence of siRNA-1: 5'-CUGACAAAGUGCCGUCCAA dTdT-3' (SEQ ID NO: 4);

[0046] Reverse sequence of siRNA-1: 5'-UUGGACGGCACUUUGUCAG TdTd-3' (SEQ ID NO: 5);

[0047] Forward sequence of siRNA-2: 5'-GAGGCAAGACGAAGAAAGU dTdT-3' (SEQ ID NO: 6);

[0048] Reverse sequence of siRNA-2: 5'-ACUUUCUUCGUCUUGCCUC TdTd-3' (SEQ ID NO: 7).

[0049] In the above two MLL-R leukemia cell lines, LAMP5-AS1 was knocked out respectively, and the cell proliferation was detected by CCK-8 assay. Experimental results such as f...

Embodiment 3

[0059] Example 3 LAMP5-AS1 targets MLL fusion protein

[0060]Whether at the cellular level or in vitro mouse experiments, LAMP5-AS1 significantly affected the function of MLL fusion gene leukemia cells. So, can this lncRNA directly regulate the expression of MLL fusion protein and thus participate in this type of disease? To investigate this question, the expression levels of MLL fusion proteins were examined by inhibiting LAMP5-AS1 by RNA interference in THP1 and MV4-11 cells. The experimental results found that knocking down LAMP5-AS1 in the MLL cell line, the protein levels of MLL-AF9 and MLL-AF4 were significantly reduced, but the mRNA of the MLL fusion protein had no significant effect ( Figure 4 A and B), suggesting that LAMP5-AS1 probably plays a role by mediating the stability or degradation of MLL fusion protein. On the other hand, in the tumor samples of NOD-SCID mice subcutaneously inoculated with shRNA Molm13, we also found that knocking down LAMP5-AS1 signific...

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Abstract

The invention discloses an application of long-chain non-coding RNA LAMP5-AS1 to MLL-R leukemia. Specifically, LAMP5-AS1 in a patient suffering from MLL fusion gene leukemia is notably and highly expressed, and besides, highly expressed LAMP5-AS1 presents notable negative correlation with the survival rate of the patient. The result of a mouse leukemia model indicates that through knocking down ofthe expression of the LAMP5-AS1, survival of MLL fusion gene leukemia can be notably restrained, and the life cycle of the model mouse can be notably prolonged. The experiment indicates that the LAMP5-AS1 can be used as a diagnosis marker and/or a treatment target point to exert effects in MLL-R leukemia.

Description

technical field [0001] The invention relates to the technical field of biomedicine, more specifically, the application of long-chain non-coding RNA LAMP5-AS1 in MLL-R leukemia. Background technique [0002] MLL fusion gene leukemia, that is, MLL (mixed-lineage leukemia) gene rearranged leukemia (MLL-rearranged leukemia, MLL-R leukemia), under the existing treatment methods, the five-year survival rate is lower than 30%. Due to the characteristics of low cure rate, poor prognosis and easy relapse in the early stage of treatment, the treatment of this type of leukemia still faces severe challenges, so it has received extensive attention in recent years. Because the five-year survival rate of MLL-R leukemia patients is significantly lower than that of other leukemia patients, many patients give up the chance of treatment. Therefore, it is necessary to adopt a new method to examine the biological characteristics of this type of leukemia, and to study the mechanism of MLL-R leuke...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886A61K45/00A61K31/713A61P35/02
CPCC12Q1/6886A61K45/00A61K31/713A61P35/02C12Q2600/158C12Q2600/178C12Q2600/136
Inventor 王文涛陈月琴孙雨蒙罗学群
Owner SUN YAT SEN UNIV
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