Application of small peptide PEARL in diagnosis and targeted therapy of acute myelogenous leukemia

Abstract

The invention belongs to the technical field of biological medicine, and particularly relates to application of small peptide PEARL in diagnosis and targeted therapy of acute myelogenous leukemia. When leukemia patient samples and leukemia cell lines are detected, obvious low expression of the small peptide PEARL in acute myelogenous leukemia patients is found. Meanwhile, the research finds that PROSER2-AS1 does not play a role, but plays a role through the coded small peptide PEARL. A mouse leukemia model experiment further finds that the overexpressed small peptide PEARL can significantly inhibit the survival of acute myelogenous leukemia cells and significantly prolong the life cycle of a model mouse. Finally, through mechanism research, it is found that the endoplasmic reticulum pressure of acute myelogenous leukemia cells is remarkably increased through overexpression of PEARL, the UPR reaction is activated, and therefore the functions of the acute myelogenous leukemia cells are affected. Therefore, the invention can provide a new accurate judgment or treatment strategy for diagnosis or targeted treatment of acute myelogenous leukemia.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and specifically relates to the application of a small peptide PEARL in the diagnosis and targeted treatment of acute myeloid leukemia. Background Art

[0002] Acute myeloid leukemia (AML) is a type of malignant tumor characterized by blocked differentiation and malignant proliferation of hematopoietic stem / progenitor cells, accounting for about 70% of all acute leukemias. At present, the pathogenesis of AML is still unclear, and the problem of drug-resistant clones frequently occurs, resulting in difficult treatment, poor prognosis and survival rate.

[0003] Therefore, the study of the same or different targets in different types of acute myeloid leukemia will play a vital role in the treatment of leukemia. Studies have shown that the unfolded protein response (UPR) is one of the main adaptive cell stress responses that leads to the development of drug and chemotherapy resistance in AML leukemia. Therefore, targeting UPR may be a new AML treatment strategy.

[0004] With the development and maturity of technologies such as transcriptomics and proteomics, more and more studies have shown that some RNA regions, including long non-coding RNA, which are traditionally considered to be non-protein-coding, actually have open reading frames (ORFs) usually less than 300 nt (nucleotides), which are called small open reading frames (sORFs), which are translated into small peptides. Transcriptome sequencing results show that a large number of non-coding RNAs (such as lncRNAs) encoding small peptides are abnormally expressed in cancer. Current studies have shown that many small peptides are dysregulated in cancer and regulate the growth, invasion and metastasis of cancer cells. In recent years, with the development of ribosome profiling technology, it has gradually been revealed that new small peptides encoded by long non-coding RNA play an important role in life activities, diseases and cancer. However, the research on small peptides is still in its infancy, and the functions of many small peptides have not yet been explored, and there is a lack of relevant research in leukemia. Summary of the invention

[0005] In order to overcome the deficiencies of the above-mentioned prior art, the present invention discovered and identified a small peptide PEARL (A Peptide located in Endoplasmic Reticulum derived from aLncRNA PROSER2-AS1) that is lowly expressed in acute myeloid leukemia, and revealed that the small peptide is an important regulatory molecule in leukemia. On the one hand, it enriches the research on small peptides, and on the other hand, it provides a theoretical basis for a comprehensive understanding of the occurrence and development of leukemia, and at the same time provides a new target for the development of drugs for acute leukemia. It can also provide a positive reference for the study of the mechanism of action of small peptides in other leukemia cancers and as a potential therapeutic target.

[0006] In order to achieve the above object, the technical solution adopted by the present invention is:

[0007] The first aspect of the present invention provides the use of a reagent for detecting the expression level of a small peptide PEARL encoded by lncRNAPROSER2-AS1 in the preparation of an acute myeloid leukemia diagnostic product, wherein the nucleic acid sequence of the lncRNAPROSER2-AS1 is shown in SEQ ID NO: 1, and the amino acid sequence of the small peptide PEARL is shown in SEQ ID NO: 2.

[0008] The present invention discovered and identified a gene encoding a small peptide PEARL, which is lowly expressed in acute myeloid leukemia but relatively highly expressed under normal physiological conditions and in other types of cancers, and the gene was named PROSER2-AS1 (ENSG00000225778). Further studies have found that in patients with acute myeloid leukemia, the expression level of PROSER2-AS1 / PEARL is significantly low, indicating that PROSER2-AS1 / PEARL has a potential clinical role as a classifier for acute myeloid leukemia.

[0009] Preferably, the reagent for detecting the expression level of the small peptide PEARL is a primer for detecting the expression amount of the small peptide PEARL, and its sequence is shown in SEQ ID NO: 3-4.

[0010] Preferably, the diagnostic product comprises a diagnostic chip or a kit.

[0011] The second aspect of the present invention provides a reagent for overexpressing a small peptide PEARL encoded by lncRNAPROSER2-AS1 in any of the following aspects 1) to 4), wherein the nucleic acid sequence of the lncRNAPROSER2-AS1 is shown in SEQ ID NO: 1, and the amino acid sequence of the small peptide PEARL is shown in SEQ ID NO: 2:

[0012] 1) Preparation of products for the treatment of acute myeloid leukemia;

[0013] 2) Preparation of products that inhibit the occurrence and development of acute myeloid leukemia;

[0014] 3) Preparation of products for inhibiting the growth of acute myeloid leukemia cells;

[0015] 4) Prepare products that promote differentiation of acute myeloid leukemia cells.

[0016] The present invention has been studied and confirmed that after overexpressing PROSER2-AS1 / PEARL using a CMV-HA tag vector, the gene functions as a translated small peptide PEARL, rather than as lncRNA PROSER2-AS1, and the small peptide can significantly inhibit the proliferation of acute myeloid leukemia cells and induce their differentiation. Moreover, experiments on the NOD-SCID mouse model show that acute myeloid leukemia cells overexpressing PEARL have the effect of inhibiting tumor growth.

[0017] Preferably, the inhibition of the occurrence and development of acute myeloid leukemia is to inhibit the translation level of acute myeloid leukemia cells.

[0018] More preferably, the inhibition of translation level of acute myeloid leukemia cells significantly increases the endoplasmic reticulum pressure of acute myeloid leukemia cells by interacting with the UPR response effector PERK, thereby activating the UPR response.

[0019] The present invention further confirms through relevant molecular biology experiments that PEARL regulates the UPR response by combining with PERK, directly affecting the endoplasmic reticulum stress of cells, specifically by increasing the endoplasmic reticulum pressure of acute myeloid leukemia cells, activating the UPR response, thereby inducing cell death and increasing the survival cycle of the disease. This shows that the present invention can regulate the expression level of PROSER2-AS1 / PEARL by genetic engineering means, thereby increasing the endoplasmic reticulum pressure of cells and inducing the UPR response, which is of great value for the precise treatment of targeted acute myeloid leukemia. At the same time, the present invention also confirms that PROSER2-AS1 / PEARL has potential clinical practical value in indicating the classification of acute myeloid leukemia.

[0020] Preferably, the reagent for overexpressing the small peptide PEARL encoded by lncRNA PROSER2-AS1 includes pCDH-CMV-MCS-EF1-Puro.

[0021] Preferably, the acute myeloid leukemia cells include MOLM-13 and HL60.

[0022] Preferably, the product is a medicine.

[0023] The third aspect of the present invention provides a therapeutic drug for acute myeloid leukemia, the drug comprising an agent for overexpressing a small peptide PEARL encoded by lncRNAPROSER2-AS1, the nucleic acid sequence of the lncRNAPROSER2-AS1 is shown in SEQ ID NO: 1, and the amino acid sequence of the small peptide PEARL is shown in SEQ ID NO: 2.

[0024] Preferably, the drug further comprises a pharmaceutically acceptable excipient.

[0025] More preferably, the excipients are functional pharmaceutical excipients available in the pharmaceutical field, including (but not limited to) surfactants, suspending agents, emulsifiers and some new pharmaceutical polymer materials, such as cyclodextrin, chitosan, polylactic acid (PLA), polyglycolic acid-polylactic acid copolymer (PLGA), hyaluronic acid, etc.

[0026] Preferably, the dosage form of the drug includes injection, powder, granule, capsule, tablet.

[0027] Preferably, the administration of the drug includes injection and oral administration.

[0028] Compared with the prior art, the present invention has the following beneficial effects:

[0029] At present, the treatment of acute myeloid leukemia at home and abroad still faces severe challenges, and there has been no report on the study of small peptides participating in the UPR response of acute myeloid leukemia. The present invention first discovered and confirmed that a small peptide PEARL encoded by a long non-coding RNA PROSER2-AS1 has a diagnostic and targeted therapeutic effect in acute myelogenous leukemia (AML). The present invention first detected leukemia patient samples and leukemia cell lines by qRT-PCR technology, and found that the small peptide PEARL was significantly low in acute myeloid leukemia patients, while it was relatively highly expressed in normal physiological conditions and other types of cancers, suggesting that the small peptide can indicate the diagnosis and prognosis of the disease. At the same time, it was found that PROSER2-AS1 itself does not play a role, but functions through the encoded small peptide PEARL. Further experiments on mouse leukemia models found that overexpression of the small peptide PEARL can significantly inhibit the survival of acute myeloid leukemia cells and significantly prolong the survival period of model mice. Finally, through the study of the biological molecular mechanism of the small peptide PEARL, it was found that overexpression of PEARL can increase the endoplasmic reticulum pressure of acute myeloid leukemia cells, activate the unfolded protein response (UPR), and directly regulate the stress level of the endoplasmic reticulum of acute myeloid leukemia cells by interacting with the UPR response effector PERK, thereby affecting the function of acute myeloid leukemia cells. Therefore, the present invention can provide a new accurate judgment or treatment strategy and genetic resources for the diagnosis or targeted treatment of acute myeloid leukemia, which has important theoretical significance and application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 Identification and localization of the small peptide PEARL encoded by PROSER2-AS1; (A) The peptide detected by mass spectrometry combined with ORFfinder analysis showed that PROSER2-AS1 ORF32 can encode a small peptide PEARL with a size of 74 amino acids starting from the non-classical start codon ACG, and the red font is the characteristic peptide of the small peptide; (B) Identification of the translation ability of PEARL, constructing HA-tagged small peptides with the start codon ACG mutated to ACA and normal small peptides respectively; (C) Antigenic epitopes prepared by polyclonal antibodies against the small peptide PEARL; (D) Detection of HA-tagged proteins using the prepared antibodies; (E) Nuclear-cytoplasmic separation showed that the small peptide PEARL was localized in the cytoplasm; (F) Immunofluorescence results showed that the small peptide PEARL was localized on the endoplasmic reticulum.

[0031] Figure 2PROSER2-AS1 / PEARL is expressed in acute myeloid leukemia; (A) GEPIA analysis showed that PROSER2-AS1 / PEARL was specifically lowly expressed in acute myeloid leukemia; (B) qRT-PCR technology detected that the expression level of PROSER2-AS1 / PEARL in the newly diagnosed samples of acute myeloid leukemia (n=50) was significantly higher than that in the normal cord blood sample group (n=10) (****P<0.0001); (C) qRT-PCR technology detected that PROSER2-AS1 / PEARL was generally lowly expressed in acute myeloid leukemia cell lines.

[0032] Figure 3 The knockdown and overexpression effects of PEARL in acute myeloid leukemia; (A) qRT-PCR technology was used to detect the knockdown effect of PEARL in HL-60; asterisks indicate that the difference between the two groups was statistically significant by t-test (*P<0.05; **P<0.01); (B) After constructing a stable PEARL overexpressing cell line, qRT-PCR and western blot techniques were used to detect the overexpression of the HA-tagged small peptide with the start codon ACG mutated to ACA and normal PEARL, respectively (***P<0.001; ****P<0.0001).

[0033] Figure 4 PEARL, rather than PROSER2-AS1, regulates the cytological function of acute myeloid leukemia; (A) overexpression of HA-tagged PEARL significantly promoted MOLM-13 differentiation; whereas overexpression of the HA-tagged small peptide PEARLMut, in which the start codon ACG was mutated to ACA, did not affect MOLM-13 cells (**P<0.01; ***P<0.001); (B) knockdown of PROSER2-AS1 / PEARL significantly inhibited MOLM-13 cell apoptosis and cell differentiation (**P<0.01; ***P<0.001).

[0034] Figure 5The animal model of acute myeloid leukemia regulated by PEARL; (A) The survival curve was used to calculate the survival period of mice after inoculation with MOLM-13 cells overexpressing PEARL; the p value was calculated using the log-rank (Mantel-Cox) test statistical method (**, p<0.01); (B) The mouse tail vein model showed that after inoculation with MOLM-13 cells overexpressing PEARL, the spleen weight of mice was significantly higher than that of the control group inoculated with NC cells (**, p<0.01); (C) The mouse tail vein model showed that after inoculation with MOLM-13 cells overexpressing PEARL, the number of cells in the mouse organs (including bone marrow, blood, spleen and liver) was significantly lower than that of the control group inoculated with NC cells; PBS injection was used as the blank control; the mean ± standard deviation of three repetitions was used in this figure, and the asterisk indicates that the difference between the two groups was statistically significant by t-test (**, p<0.01; ***, p<0.001).

[0035] Figure 6 PEARL regulates cellular endoplasmic reticulum stress; (A) SUnSET experiments showed that overexpression of PEARL inhibited puromycin insertion, indicating that the overall translation level of the protein was reduced; (B) GO analysis of PEARL-interacting proteins showed that PEARL was related to the unfolded protein response; (C) Immunoprecipitation experiments confirmed that PEARL was tightly bound to the UPR response effector PERK. DETAILED DESCRIPTION

[0036] The specific embodiments of the present invention are further described below. It should be noted that the description of these embodiments is used to help understand the present invention, but does not constitute a limitation of the present invention. In addition, the technical features involved in each embodiment of the present invention described below can be combined with each other as long as they do not conflict with each other.

[0037] The experimental methods in the following examples are conventional methods unless otherwise specified, and the experimental materials used in the following examples are commercially available unless otherwise specified.

[0038] Example 1: Identification and localization of the small peptide PEARL encoded by PROSER2-AS1

[0039] The present invention discovered a lncRNA, which was named PROSER2-AS1 (ENSG00000225778, NR_038222.1). The gene locus of PROSER2-AS1 (reference hg38) is located on the antisense DNA strand of human chromosome 10. Its gene covers a range from 11,847,213 bp to 11,894,710 bp and can transcribe a lncRNA with a length of 3163 nt. Its nucleotide sequence is shown in SEQ ID NO: 1. At the same time, the present invention newly identified that the lncRNA has a short peptide PEARL ( Figure 1 A), the sequence of which is shown in SEQ ID: 2.

[0040] PROSER2-AS1 gene sequence (3163 bp, SEQ ID NO: 1):

[0041] >PROSER2-AS1(ENSG00000225778,NR_038222.1):

[0042]

[0043] PEARL amino acid sequence (74aa, SEQ ID NO: 2):

[0044] >PEARL

[0045] TQQKGQDTQRRWRTHLGTEGQCDLPGAGGPARAFPEETAKPRPGTAEEKQGGGRRGPVPSSSAVPGGRSARLSA.

[0046] In order to further verify the existence of the small peptide, primers (SEQ ID NO: 3 and SEQ ID NO: 4) were designed to amplify the small peptide, and the small peptide was verified by western blot, and it was localized using immunofluorescence technology and confocal microscopy. In this example, an HA tag was added to the C-terminus of ORF32 (chr10: 11925853-11937442 strand = -). Figure 1 B).

[0047] Forward primer sequence: 5'ACGCAGCAGAAGGGACAAGAC 3' (SEQ ID NO: 3);

[0048] Reverse primer sequence: 5'CGGAATTCTATGTTTGCTTTGTTTTATTAACTCCTTGGAG 3' (SEQ ID NO: 4);

[0049] At the same time, a plasmid with the start codon ACG mutated to ACA was constructed as a control, and primers (SEQ ID NO:5 and SEQ ID NO:6) were designed for amplification and verified by western blot. Western blot results showed that ORF32 of PROSER2-AS1 could translate the small peptide PEARL, but when ACG was mutated to ACA, the small peptide could not be expressed.

[0050] Forward primer sequence: 5'ACACAGCAGAAGGGACAAGAC 3' (SEQ ID NO: 5);

[0051] Reverse primer sequence: 5'CGGAATTCTATGTTTGCTTTGTTTTATTAACTCCTTGGAG 3' (SEQ ID NO: 6).

[0052] At the same time, in order to further identify the endogenous expression of the small peptide PEARL, an endogenous antibody was customized based on the characteristic peptide segment of the small peptide ( Figure 1C), western blot results show that the antibody can detect the corresponding trend changes of the small peptide when the small peptide is overexpressed using pCDH-CMV-MCS-EF1-Puro (CD510B-1) ( Figure 1 D), these results not only show that the customized antibody is specific, but also show that the small peptide encoded by PROSER2-AS1 actually exists in vivo. Using nuclear cytoplasm separation experiments, it was found that the small peptide PEARL is mainly located in the cytoplasm ( Figure 1 E). Finally, immunofluorescence experiments revealed that the small peptide PEARL was localized on the endoplasmic reticulum ( Figure 1 F).

[0053] Example 2: PEARL expression analysis and clinical value assessment

[0054] The preliminary study of the present invention found that the small peptide PEARL is specifically low expressed in acute myeloid leukemia, but highly expressed in normal physiological conditions and other types of leukemia ( Figure 2 A and C). In order to further determine the expression specificity of PEARL in acute myeloid leukemia, this example collected a batch of patient bone marrow samples from the First Affiliated Hospital of Sun Yat-sen University for detection and analysis, including 50 newly diagnosed samples of acute myeloid leukemia and 10 normal cord blood samples. All sample collections were approved by the Ethics Committee of Sun Yat-sen University and informed consent was obtained from the patients. PEARL was specifically detected by extracting RNA and using qRT-PCR technology. The qRT-PCR primers used in this process are as follows:

[0055] Forward primer sequence: 5'CTGGTGGCCTCCTGCTTAC 3' (SEQ ID NO: 7);

[0056] Reverse primer sequence: 5'CGTGTCTTGTCCCTTCTGCT 3' (SEQ ID NO: 8).

[0057] The experiment found that PEARL was significantly lower expressed in acute myeloid leukemia compared with normal cord blood samples (p<0.001) ( Figure 2 B), while relatively high expression in normal physiological conditions and other types of cancer ( Figure 2 C). This indicates that the expression level of PEARL may be related to the incidence of the disease and can indicate the prognosis of acute myeloid leukemia. The above results indicate that PEARL has the potential to distinguish acute myeloid leukemia from normal samples; and to a certain extent indicates the prognosis of the disease, suggesting that overexpression of PEARL may be used as a treatment for acute myeloid leukemia.

[0058] Example 3: Functional identification of PEARL in acute myeloid leukemia

[0059] In order to solve the core problem of PEARL's involvement in the regulation of acute myeloid leukemia, this example intends to further study the effect of PEARL on the function of acute myeloid leukemia. First, siRNA was used to knock down the small peptide PEARL, and RT-PCR technology was used to detect the knockdown effect of PEARL in HL-60 ( Figure 3 A); After constructing a stable PEARL overexpressing cell line, qRT-PCR and western blot were used to detect the overexpressed HA-tagged small peptide with the start codon ACG mutated to ACA and normal PEARL ( Figure 3 B). Identify whether it is the small peptide PEARL or the lncRNA PROSER2-AS1 that performs the function. Through experimental research, overexpression of HA-tagged PEARL can significantly promote the differentiation of MOLM-13 cells; while overexpression of the small peptide PEARLMut with the start codon ACG mutated to ACA of the HA tag has no effect on MOLM-13 cells ( Figure 4 A), which indicates that PEARL functions in the form of protein, not RNA. In addition, two different siRNA sequences were designed for PEARL using siRNA interference technology, as shown below:

[0060] Forward sequence of siRNA-1: 5'GGUUGACCUGAGCCUACUU dTdT 3' (SEQ ID NO: 9);

[0061] Reverse sequence of siRNA-1: 5'AAGUAGGCUCAGGUCAACC dTdT3' (SEQ ID NO: 10);

[0062] Forward sequence of siRNA-2: 5'GCGGAUCGAGGACUGCCUA dTdT 3' (SEQ ID NO: 11);

[0063] Reverse sequence of siRNA-2: 3'UAGGCAGUCCUCGAUCCGC dTdT5' (SEQ ID NO: 12).

[0064] In the acute myeloid leukemia cell line MOLM-13, after knocking down PEARL, flow cytometry was used to detect cell apoptosis. Figure 4 As shown in B, when PEARL was knocked down, the apoptosis rate of cells was significantly reduced. At the same time, after knocking down PEARL by siRNA interference technology, flow cytometry was used to detect the differentiation of cells. Figure 4C shows that when PEARL is knocked down, the differentiation of acute myeloid leukemia cell lines is significantly inhibited. Therefore, it is speculated that PEARL has a potential regulatory effect on the occurrence and development of acute myeloid leukemia.

[0065] Next, the regulatory effect of PEARL on acute myeloid leukemia was further verified at the adult level. The pCDH-CMV-MCS-EF1-Puro (CD510B-1) lentiviral expression system was also used to construct a stable overexpression strain of PEARL, and the MOLM-13PEARL overexpression cell line was obtained by puromycin screening. Subsequently, the validated stable MOLM-13PEARL cell line was expanded and cultured, and inoculated into 5-week-old NOD-SCID mice by tail vein injection: 2 groups (NC, PEARL), 10 mice in each group, and PBS was injected as a blank control (each mouse was inoculated with 1×10 6 The animal experiment was approved by the Animal Ethics Committee of Sun Yat-sen University. The survival curve analysis of the Log-rank (Mantel-Cox) Test showed that the survival rate of the mice inoculated with PEARL overexpression was higher than that of the control group ( Figure 5 A). Three weeks after inoculation, the bone marrow, peripheral blood and organs of each group of mice were collected. The spleen size and weight of the PEARL overexpressing cell group were significantly higher than those of the NC cell inoculation group ( Figure 5 B). At the same time, human hCD45 antibody was used to mark the surface of human leukemia cells and detect them. The flow cytometry results showed that CD45-positive MOLM-13 cells were basically undetectable in the PBS group, and the number of CD45-positive MOLM-13 cells in various organs of the PEARL group mice was significantly less than that of the NC group mice ( Figure 5 C) The above results indicate that overexpression of PEARL can inhibit the tumorigenesis of acute myeloid leukemia and affect the growth of such leukemia cells, thereby affecting the occurrence and development of acute myeloid leukemia. It also indicates that PEARL may be a potential target for the treatment of acute myeloid leukemia.

[0066] Example 4: PEARL binds to PERK to regulate endoplasmic reticulum stress levels in acute myeloid leukemia cells

[0067] Both at the cellular level and in vitro mouse experiments have confirmed that PEARL can significantly affect the function of acute myeloid leukemia cells. So how does PEARL specifically regulate acute myeloid leukemia? To solve this problem, this example uses SUnSET experiments to find that after overexpressing PEARL in MOLM-13 cells, the puromycin-labeled peptide segment is significantly reduced, indicating that overexpression of PEARL can inhibit the translation level of acute myeloid leukemia cells ( Figure 6A). Further analysis of PEARL interacting proteins revealed that PEARL is associated with the unfolded protein response ( Figure 6 B). Finally, immunoprecipitation was used to identify that PEARL can specifically interact with the UPR effector PERK ( Figure 6 C). The above results indicate that after overexpression of PEARL, PEARL directly regulates the stress level of the endoplasmic reticulum of acute myeloid leukemia cells by interacting with the UPR response effector PERK, significantly increasing the endoplasmic reticulum pressure of acute myeloid leukemia cells and activating the UPR response, thereby inhibiting the translation level of acute myeloid leukemia cells and affecting the function of acute myeloid leukemia cells, ultimately achieving the effect of affecting the occurrence and development of acute myeloid leukemia.

[0068] In summary, the present invention has discovered and confirmed for the first time that a small peptide PEARL based on lncRNA translation is lowly expressed in acute myeloid leukemia, but relatively highly expressed in other types of cancers under normal physiological conditions, suggesting that the small peptide can indicate the diagnosis and prognosis of the disease. In addition, the present invention also illustrates that PEARL regulates the endoplasmic reticulum stress level of acute myeloid leukemia, and overexpression of PROSER2-AS1 / PEARL can significantly increase the endoplasmic reticulum pressure of acute myeloid leukemia cells, activate the UPR response, thereby inducing cell death, and increasing the survival cycle of the disease. These findings can provide a new theoretical basis for the development of drug targets for acute myeloid leukemia, and have important application value for the diagnosis and gene-targeted treatment of acute myeloid leukemia.

[0069] The embodiments of the present invention are described in detail above, but the present invention is not limited to the described embodiments. For those skilled in the art, various changes, modifications, substitutions and variations of these embodiments are made without departing from the principles and spirit of the present invention, and still fall within the protection scope of the present invention.

Claims

1. Use of a reagent for detecting the expression level of a small peptide PEARL encoded by lncRNA PROSER2-AS1 in the preparation of a diagnostic product for acute myeloid leukemia, characterized in that: The nucleic acid sequence of the lncRNAPROSER2-AS1 is shown in SEQ ID NO: 1, and the amino acid sequence of the small peptide PEARL is shown in SEQ ID NO:

2.

2. The use according to claim 1, characterized in that: The reagent for detecting the expression level of the small peptide PEARL is a primer for detecting the expression amount of the small peptide PEARL, and its sequence is shown in SEQ ID NO: 3-4.

3. The use according to claim 1, characterized in that: The diagnostic product includes a diagnostic chip or a kit.

4. Use of a reagent for overexpressing a small peptide PEARL encoded by lncRNA PROSER2-AS1 in any of the following aspects 1) to 4), characterized in that: The nucleic acid sequence of the lncRNAPROSER2-AS1 is shown in SEQ ID NO: 1, and the amino acid sequence of the small peptide PEARL is shown in SEQ ID NO: 2: 1) Preparation of products for the treatment of acute myeloid leukemia; 2) Preparation of products that inhibit the occurrence and development of acute myeloid leukemia; 3) Preparation of products for inhibiting the growth of acute myeloid leukemia cells; 4) Prepare products that promote differentiation of acute myeloid leukemia cells.

5. The use according to claim 4, characterized in that: The method of inhibiting the occurrence and development of acute myeloid leukemia is to inhibit the translation level of acute myeloid leukemia cells.

6. The use according to claim 5, characterized in that: The inhibition of translation level of acute myeloid leukemia cells significantly increases the endoplasmic reticulum pressure of acute myeloid leukemia cells by interacting with the UPR response effector PERK, thereby activating the UPR response.

7. The use according to claim 4, characterized in that: The reagent for overexpressing the small peptide PEARL encoded by lncRNA PROSER2-AS1 includes pCDH-CMV-MCS-EF1-Puro.

8. The use according to claim 4, characterized in that: The acute myeloid leukemia cells include MOLM-13 and HL60.

9. The use according to claim 4, characterized in that: The product described is a drug.

10. A therapeutic drug for acute myeloid leukemia, characterized in that: The drug includes an agent for overexpressing a small peptide PEARL encoded by lncRNA PROSER2-AS1, the nucleic acid sequence of the lncRNA PROSER2-AS1 is shown as SEQ ID NO: 1, and the amino acid sequence of the small peptide PEARL is shown as SEQ ID NO: 2.

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