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Application of IGSF6 to preparing multiple myeloma diagnosis and treatment products

A multiple myeloma, product technology, applied in the field of tumor diagnosis, treatment, and prognosis prediction, can solve problems such as incurable

Active Publication Date: 2016-12-14
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although with the in-depth study of the pathogenesis of MM, some research results and new treatment options have made significant progress in the treatment of MM, so far, MM is still an incurable disease.

Method used

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  • Application of IGSF6 to preparing multiple myeloma diagnosis and treatment products
  • Application of IGSF6 to preparing multiple myeloma diagnosis and treatment products
  • Application of IGSF6 to preparing multiple myeloma diagnosis and treatment products

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 the gene that has not been studied in the prior art on the correlation between this gene and multiple myeloma as a candidate gene, and considering the results of gene sequencing, select the IGSF6 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 transcriptase 1 μL...

Embodiment 3I

[0120] Embodiment 3 IGSF6 gene overexpression

[0121] 1. Plasmid construction

[0122] Amplification primers are designed according to the coding sequence of the IGSF6 gene, and the design of the primers is well known to those skilled in the art. Amplify the coding sequence of the full-length IGSF6 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 -IGSF6 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...

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Abstract

The invention discloses a diagnosis and treatment target-IGSF6 gene for the multiple myeloma. By detecting the content of the IGSF6 gene and expression products of the IGSF6 gene in myeloid tissue of a testee, whether the testee suffers from the multiple myeloma or not can be judged or whether the testee bears a risk of suffering from the multiple myeloma or not can be diagnosed. In addition, by studying the proliferation, migration and invasion indexes of in-vitro cultured cells of the multiple myeloma, it is proved that the IGSF6 gene serves as a medicine target for treating 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 IGSF6 abnormality; and a tumor therapeutic agent for activating IGSF6 gene or protein. Background technique [0002] Multiple myeloma (MM) is a malignant proliferation of plasma cells characterized by abnormal proliferation of bone marrow plasma cells accompanied by excessive production of monoclonal immunoglobulins or light chains. The clinical manifestations mainly include osteolytic injury, anemia, hypercalcemia, kidney injury and repeated infection. Although the research on the pathogenesis of MM has deepened, some research results and new treatment options have made significant progress in the treatment of MM, but so far, MM is still an incurable disease. Therefore, it is still a huge task for clinicians and researchers t...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/68G01N33/574A61K45/00A61P35/00
Inventor 杨承刚肖枫边洋
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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