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Application of miR-145-3p in preparation of apoptosis and autophagy enhancer

An enhancer, cell technology, applied in the field of clinical application of miR-145-3p and its analogs, to achieve the effect of promoting cell apoptosis

Active Publication Date: 2019-05-24
SECOND AFFILIATED HOSPITAL SECOND MILITARY MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

There is also no literature report on the relationship between miR-145-3p and multiple myeloma

Method used

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  • Application of miR-145-3p in preparation of apoptosis and autophagy enhancer
  • Application of miR-145-3p in preparation of apoptosis and autophagy enhancer
  • Application of miR-145-3p in preparation of apoptosis and autophagy enhancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] Example 1, microRNAs expression analysis and verification in MM patients

[0076] There are many kinds of microRNAs in the peripheral plasma of MM patients. In this experiment, the classic microarray screening method was used to screen the microRNAs differentially expressed in MM patients and normal controls.

[0077] The criteria for our selection of patients are as follows: all 30 patients with multiple myeloma were inpatients in Shanghai Changzheng Hospital (21 males and 9 females, aged 40-74 years, median 60 years old, ISS stage 2 stage and stage 3 patients), and the hospitalization time was from February 2013 to July 2014. These patients all met the diagnostic criteria for multiple myeloma recommended by NCCN in 2013: (1) tissue biopsy or bone marrow smear: plasma cells>30 %, often accompanied by morphological changes; (2) Monoclonal immunoglobulin (M protein): IgG>35g / L, IgA>20g / L, IgM>15g / L, IgD>2g / L, IgE>2g / L , monoclonal K or lambda light chain in urine > 1g / ...

Embodiment 2

[0084] Example 2, the value of miR-145-3p in clinical parameters and prognosis

[0085] Serum collection from patients and detection of clinical parameters: After collecting serum from MM patients in a sampling tube without anticoagulant, leave it for a period of time, centrifuge at 3500 rpm for 10 minutes, and collect the supernatant for later use. For the detection of various clinical indicators, albumin is detected by BCG colorimetric method, β2-MG is detected by immunoturbidimetric method, creatinine is detected by colorimetric method, Ca 2+ Visible absorbance photometry method was used for detection.

[0086] Collection of patient clinical prognosis information: Obtain patient survival information through follow-up and other methods.

[0087] Statistical methods: We used correlation analysis and Kaplan-Meier survival analysis to analyze the clinical application value of miR-145-3p.

[0088] The result is as figure 2 As shown, the correlation analysis results between t...

Embodiment 3

[0089] Example 3, miR-145-3p overexpression induces increased cell apoptosis

[0090] MM cell culture: Human myeloma cell line LP-1 (purchased from the Institute of Cells, Chinese Academy of Sciences) was cultured under conventional conditions, that is, in a 37°C, 5% carbon dioxide incubator, and the medium contained 10% fetal bovine serum (FBS) and 1% Double antibody (penicillin 100U / ml and streptomycin 0.1mg / ml) 1640.

[0091] MiR-145-3p analogs and their inhibitors were designed and synthesized by Guangzhou Ruibo Biotechnology Co., Ltd., and then transfected:

[0092] miR-145-3p mimic, the sequence is shown in SEQ ID NO:1;

[0093] miR-145-3p inhibitor, the sequence is 5'-AGAACAGUAUUUCCAGGAAUCC-3' (SEQ ID NO: 2);

[0094] Adjust the LP-1 cells to the best state, adjust the concentration after counting, and the cell density is 1x10 5 / ml; pipette 2.5μl miR-145-3p mimic / inhibitors into 100ul Opti-MEM, and mix gently. At the same time, set up a transfection control to ensu...

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Abstract

The invention relates to the technical field of biomedicine. Analysis of microRNAs expression profile is carried out in peripheral plasma of a patient with multiple myeloma; verifying is performed bymeans of qPCR (quantitative polymerase chain reaction); the expression level of miR-145-3p in the peripheral plasma of the patient with multiple myeloma is evidently lower than that of normal healthypeople. The results from the observation of apoptosis and autophagy level through miR-145-3p overexpression and miR-145-3p silencing show that miR-145-3p can promote apoptosis and autophagy. A novel serological marker is provided for the diagnosis of multiple myeloma; a novel target is also provided for the prevention and treatment of multiple myeloma.

Description

[0001] This application is a divisional application of the invention patent application submitted on September 23, 2016 with the patent application number 201610845836.0 and the invention name "miR-145-3p in the preparation of drugs for the prevention or treatment of multiple myeloma". technical field [0002] The present invention relates to the technical field of biomedicine, more specifically, relates to the clinical application of miR-145-3p and its analogues. Background technique [0003] MicroRNA is a kind of small molecule RNA with post-transcriptional regulatory activity discovered in Drosophila, nematode, mouse and human in recent years. The discovery of a large number of endogenous small RNAs benefits from two technologies, one is the construction and sequencing of microRNA cDNA libraries; the other is the capture of biotin-labeled oligonucleotide probes, which are A technique for performing PCR amplification. Through library construction and sequencing, people ha...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/7105A61P35/00A61K31/573
CPCA61K31/573A61K31/7105C12Q1/6886C12Q2600/136C12Q2600/158C12Q2600/178A61K2300/00
Inventor 周琳吴洪坤刘畅王皓殷杏吴雯李江燕
Owner SECOND AFFILIATED HOSPITAL SECOND MILITARY MEDICAL UNIV
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