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Diagnosis and treatment product for multiple myeloma biomarker

A multiple myeloma and product technology, applied in the field of tumor diagnosis, prognosis prediction, and treatment, can solve problems such as no clear answer and unclear pathogenesis of multiple myeloma

Active Publication Date: 2016-12-07
GUAN BOJIAN BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the pathogenesis of multiple myeloma is still unclear. Existing research results have shown that it is mainly related to genetic changes and the role of cytokines, but there is no clear answer so far.

Method used

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  • Diagnosis and treatment product for multiple myeloma biomarker
  • Diagnosis and treatment product for multiple myeloma biomarker
  • Diagnosis and treatment product for multiple myeloma biomarker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] Example 1 Gene Chip Screening for Differentially Expressed Genes

[0075] 1. Materials:

[0076] 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.

[0077] 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.

[0078] 2. Obtaining tissue RNA

[0079] Total tissue RNA was extracted using the Trizol one-step method.

[0080] 3. Determination of RNA purity and concentration

[0081] 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

[0094] Example 2 Large sample validation of differentially expressed genes screened out

[0095] 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 GTF2F1 gene (its expression is upregulated in multiple myeloma tissues) for verification .

[0096] 1. Sample collection

[0097] According to the method of Example 1, 50 cases of multiple myeloma tissues and 60 cases of normal bone marrow tissues were collected.

[0098] 2. Validation at the mRNA level

[0099] 2.1 Extract tissue RNA

[0100] Step is with embodiment 1.

[0101] 2.2 Reverse transcription

[0102] 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 3

[0125] Example 3 Inhibition of GTF2F1 Gene Expression

[0126] 1. siRNA design and synthesis

[0127] siRNA sequences against GTF2F1:

[0128] siRNA-GTF2F1:

[0129] The sense strand is 5'-AUGAUGUUAUAUUUUUUUGGUU-3' (SEQ ID NO.5)

[0130] The antisense strand is 5'-CCAAAAAAUAUAACAUCAUGG-3' (SEQ ID NO.6);

[0131] The above siRNA sequences and negative control siRNA sequences (siRNA-NC) were provided by Shanghai Gemma Pharmaceutical Technology Co., Ltd.

[0132] 2. Culture and transfection of multiple myeloma cells

[0133] 2.1 Cell culture

[0134] RPMI8226 cells were placed in RPMI 1640 medium containing 15% fetal bovine serum (FBS), penicillin 100 U / ml, and streptomycin 100 μg / ml at 37°C, 5% CO 2 , cultivated in a saturated humidity environment.

[0135] 2.2 Cell transfection

[0136] (1) 24 hours before transfection, inoculate 0.5-2*10 5 cells, the cell confluency was 30-50% at the time of transfection. When plating, the cells should be digested and mixed completel...

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Abstract

The invention discloses a GTF2F1 gene and an expression product thereof. The GTF2F1 gene and the expression product can be used as a molecular marker for multiple myeloma diagnosis and treatment. Whether a subject has multiple myeloma or has the risk of suffering the multiple myeloma or not can be determined by detecting the content of GTF2F1 gene and expression product thereof in myeloid tissues of the subject. Studies on proliferation, apoptosis, migration and invasion of an in-vitro cultured multiple myeloma cell discover that, by inhibiting expression of the GTF2F1 gene, cell proliferation can be inhibited, cell apoptosis can be promoted, and cell migration can be inhibited. Studying results indicate that the GTF2F1 gene and the expression product thereof are a potential medicine target for multiple myeloma treatment.

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 GTF2F1 abnormality; and a tumor therapeutic agent for inhibiting GTF2F1 gene or protein. Background technique [0002] Multiple myeloma (MM) is a malignant tumor characterized by abnormal proliferation and accumulation of monoclonal plasma cells in the bone marrow, and it is more common in the elderly over 60 years old. At present, the incidence of multiple myeloma in my country is about 1 / 100,000 to 2 / 100,000, ranking second in the incidence of hematological malignancies. With the acceleration of population aging, the incidence of multiple myeloma in my country will be on the rise. At present, the pathogenesis of multiple myeloma is still unclear. Existing research results show that it is mainly related to two aspects: geneti...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/68G01N33/574A61K45/00A61P35/00
CPCA61K45/00C12Q1/6886C12Q2600/118C12Q2600/136C12Q2600/158G01N33/57407G01N33/57484G01N33/68
Inventor 杨承刚宋宏涛
Owner GUAN BOJIAN BIOTECH CO LTD
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