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Use of ms4a6a as a marker for diagnosis and treatment of multiple myeloma

A technology for MS4A6A and multiple myeloma, which is applied in the field of tumor diagnosis, treatment, and prognosis prediction, and can solve the problems that the treatment has not been significantly improved, and the prognosis of patients has not been significantly improved.

Active Publication Date: 2019-10-11
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, despite the clinical application of thalidomide and protease inhibitors, as well as allogeneic bone marrow transplantation technology, the treatment of MM has not been significantly improved, and the prognosis of patients has not been significantly improved.

Method used

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  • Use of ms4a6a as a marker for diagnosis and treatment of multiple myeloma
  • Use of ms4a6a as a marker for diagnosis and treatment of multiple myeloma
  • Use of ms4a6a as a marker for diagnosis and treatment of multiple myeloma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Example 1 Gene Chip Screening for Differentially Expressed Genes

[0070] 1. Materials:

[0071] Multiple myeloma tissue: A total of 10 bone marrow biopsy specimens were collected from confirmed MM patients. The diagnostic criteria for MM refer to the "Chinese Guidelines for the Diagnosis and Treatment of Multiple Myeloma 2011 Revised Edition". Among the patients, there were 5 males and 5 females, with a median age of 59 years.

[0072] Normal bone marrow tissue: 10 bone marrow biopsy specimens were collected from patients with malnutrition anemia at the same period as the control group, including 5 males and 5 females, with a median age of 53 years.

[0073] 2. Obtaining tissue RNA

[0074] Total tissue RNA was extracted using the Trizol one-step method.

[0075] 3. Determination of RNA purity and concentration

[0076] Take 1 μl of RNA solution, measure OD260 and OD280 with the instrument, the RNA concentration is OD260 value × dilution factor × 40 / 1000, calculate...

Embodiment 2

[0089] Example 2 Large sample verification screened out differentially expressed genes

[0090] Considering that there is no gene in the prior art that has been studied on the correlation between this gene and multiple myeloma as a candidate gene, and considering the results of gene sequencing, select the MS4A6A gene (its expression is down-regulated in multiple myeloma tissues) for verification .

[0091] 1. Sample collection

[0092] According to the method of Example 1, 50 cases of multiple myeloma tissues and 60 cases of normal bone marrow tissues were collected.

[0093] 2. Validation at the mRNA level

[0094] 2.1 Extract tissue RNA

[0095] Step is with embodiment 1.

[0096] 2.2 Reverse transcription

[0097] A total of 20 μL of reverse transcription system, including 2 μg / 2 μL of total cell RNA, 1 μL of 50 U / μL Rnasin, 4 μL of 5× reverse transcription reaction buffer, 2 μl of 10 mM d NTP, 2 μL of 50 μg / mL random primer (promega), 200 U / μL M-MLV reverse Recording...

Embodiment 3

[0120] Example 3 MS4A6A gene overexpression

[0121] 1. Plasmid construction

[0122] Amplification primers were designed according to the coding sequence of the MS4A6A gene, and the design of primers is well known to those skilled in the art. Amplify the coding sequence of the full-length MS4A6A gene from the cDNA library of adult fetal brain (clontech company, article number: 638831), insert the above cDNA sequence into the eukaryotic cell expression vector pcDNA3.1, and connect the obtained recombinant vector pcDNA3.1 -MS4A6A was used in subsequent experiments.

[0123] 2. Culture and transfection of multiple myeloma cells

[0124] 2.1 Cell culture

[0125] RPMI8226 cells were placed in RPMI 1640 medium containing 15% fetal bovine serum (FBS), 100 U / ml penicillin, and 100 μg / ml streptomycin at 37°C, 5% CO 2 , cultivated in a saturated humidity environment.

[0126] 2.2 Cell transfection

[0127] (1) The day before transfection, 0.5-2*10 5 Tumor cells were suspended in ...

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Abstract

The invention discloses a genetic marker, namely, MS4A6A. The MS4A6A can be used for judging whether a subject has a risk of suffering from multiple myeloma or not or judging whether the subject suffers from the multiple myeloma. In additions, the MS4A6A can also be used for preparing drugs for treating the multiple myeloma. According to the MS4A6A, a novel diagnostic method is provided for diagnosing the multiple myeloma at the molecular level clinically, and meanwhile a novel drug target is provided for a gene therapy of the multiple myeloma.

Description

technical field [0001] The present invention relates to the fields of tumor diagnosis, treatment, and prognosis prediction, more specifically, the present invention relates to a method for tumor diagnosis and prognosis prediction by means of detecting MS4A6A abnormality; and a tumor therapeutic agent for activating MS4A6A gene or protein. Background technique [0002] Multiple myeloma (multiple myeloma) is a relatively common malignant tumor of the blood system. It is a disease caused by abnormal clonal proliferation of plasma cells in the body. abnormal clonal proliferation of myeloma cells). Due to abnormal plasma cells infiltrating bone, bone marrow and multiple organ tissues in the whole body, causing pathological bone destruction, abnormal monoclonal immunoglobulin appears in the patient's serum, thereby inhibiting the body's normal polyclonal immunoglobulin, and the patient's M protein appears in the urine, and the patient finally shows anemia, renal function damage a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886G01N33/68G01N33/574A61K45/00A61P35/00
Inventor 杨承刚肖枫任静
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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