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miRNAs expression profile model related to human multiple myeloma, construction method and application thereof

A multiple myeloma and atlas model technology, applied in the biological field, can solve problems such as poor compliance of elderly patients, missed diagnosis of MM, and inaccurate evaluation

Pending Publication Date: 2019-12-13
INST OF HEMATOLOGY & BLOOD DISEASES HOSPITAL CHINESE ACADEMY OF MEDICAL SCI & PEKING UNION MEDICAL COLLEGE
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  • Description
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AI Technical Summary

Problems solved by technology

[0002] At present, the most important and basic diagnostic and prognostic methods for multiple myeloma (MM) are based on bone marrow biopsy, which is traumatic, and the compliance of elderly patients is poor, and it cannot be repeated in a short period of time.
In addition, the onset of MM has the characteristics of scattered distribution of multiple lesions, and a single site of bone marrow puncture cannot reflect the overall characteristics of the patient's tumor cells, which has also become an important reason for the missed diagnosis of MM and inaccurate disease evaluation.

Method used

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  • miRNAs expression profile model related to human multiple myeloma, construction method and application thereof
  • miRNAs expression profile model related to human multiple myeloma, construction method and application thereof
  • miRNAs expression profile model related to human multiple myeloma, construction method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1: Study on differential expression profile of miRNA in tumor cells of myeloma patients

[0039] Bone marrow samples were collected from patients with multiple myeloma (n=8), mononuclear cells were separated with lymphocyte separation medium, and CD138 cells were obtained by magnetic bead sorting + Plasma cell. The RNA of plasma cells (Plasma Cells, PC) was extracted, and the miRNA expression profile was identified by next-generation small RNA sequencing. The miRNA expression profile of normal human (n=2) plasma cells was used as a control to clarify the characteristic miRNA expression of tumor cells in myeloma patients Spectrum (such as figure 1 , figure 2 Shown, among which the multiple myeloma patients are circled in the PCA graph). The differential genes obtained by sequencing are shown in Table 1 (P<0.05); there were 376 differential genes in the sequencing results, of which 293 were up-regulated and 83 were down-regulated.

[0040] Table 1. Differenti...

Embodiment 2

[0051] Example 2: Research on the expression profile of serum miRNA in patients with myeloma

[0052] Collect peripheral blood samples (Peripheral Blood Serum, PBS) from multiple myeloma patients (n=8), separate serum and extract miRNA molecules (extraction kit, QiagenLot: 160022868), and use next-generation small RNA sequencing to identify miRNA expression profiles , using the serum miRNA expression profile of normal people (n=2) as a control, to clarify the characteristic serum miRNA expression profile of myeloma patients (such as image 3 , Figure 4 Shown, among which the multiple myeloma patients are circled in the PCA graph). The analysis results showed that compared with the normal human serum miRNA expression profile, there were 74 differentially expressed miRNAs in the serum of myeloma patients (P<0.05), of which 18 differentially expressed genes were up-regulated and 56 genes were down-regulated. PBS The differential miRNAs genes are shown in Table 2.

[0053] Tab...

Embodiment 3

[0057] Example 3: Study on differential miRNA expression profile of serum exosomes in patients with multiple myeloma

[0058] Peripheral blood samples from patients with multiple myeloma (n=8) were collected, exosomes (Exosome, EXO) were isolated and purified, and total RNA molecules in the exosomes were extracted. Identification of miRNA expression profile through quality inspection, library construction, sequencing, and smallRNA sequencing, using the serum exosomal miRNA expression profile of normal people (n=2) as a control, to clarify the characteristic serum miRNA expression profile of myeloma patients (such as Figure 5 , Image 6 Shown, among which the multiple myeloma patients are circled in the PCA graph). The sequencing results showed that compared with normal human serum miRNA, there were a total of 21 differential genes (P<0.05), of which 14 differential genes were up-regulated and 7 genes were down-regulated, as shown in Table 3.

[0059] Table 3. Differential m...

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Abstract

The invention discloses a miRNA expression profile model related to human multiple myeloma, a construction method and an application thereof. The invention discloses a miRNAs expression profile of tumor cells and circulating body fluid of a patient suffering from multiple myeloma, determines a molecular map of differentially expressed miRNAs between the patient suffering from multiple myeloma andhealthy people, and screens a biological marker for early diagnosis, prognosis evaluation and curative effect prediction of myeloma diseases, analyzes the myeloma cell characteristic miRNAs molecularexpression profile, correspondingly regulates mRNA in a targeted manner, and explores the correlation between differential miRNA and mRNA molecules and myeloma disease progression and the molecular mechanism of the correlation. The miRNAs expression profile disclosed by the invention provides a theoretical basis and an experimental basis for the treatment of multiple myeloma.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a miRNA expression profile model, construction method and application related to human multiple myeloma. Background technique [0002] At present, the most important and basic diagnostic and prognostic methods for multiple myeloma (MM) are based on bone marrow biopsy, which is traumatic, poor compliance for elderly patients, and cannot be repeated in a short period of time. In addition, the onset of MM is characterized by scattered distribution of multiple lesions, and bone marrow aspiration at a single site cannot reflect the overall characteristics of tumor cells in patients, which has also become an important reason for missed diagnosis of MM and inaccurate disease evaluation. Therefore, it is of great theoretical and practical significance for the clinical diagnosis and treatment of MM to develop simple, easy, and less invasive biomarkers for early diagnosis, curative effect moni...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6869C12N15/11
CPCC12Q1/6886C12Q1/6869C12Q2600/158C12Q2600/178C12Q2535/122C12Q2545/113
Inventor 郝牧魏晓晶邱录贵
Owner INST OF HEMATOLOGY & BLOOD DISEASES HOSPITAL CHINESE ACADEMY OF MEDICAL SCI & PEKING UNION MEDICAL COLLEGE
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