Application of RPL8 to preparation of tool for diagnosing or treating multiple myeloma

A multiple myeloma, tool technology, applied in the direction of anti-tumor drugs, biochemical equipment and methods, microbial determination/examination, etc., can solve problems such as inevitable recurrence

Inactive Publication Date: 2016-10-12
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the main means of clinical treatment of various stages of MM are hormones, alkyl drugs, immunomodulators, protease inhibitors, etc. These methods can alleviate MM, but eventually relapse is inevitable, so some new diagnosis and treatment strategies need to be developed

Method used

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  • Application of RPL8 to preparation of tool for diagnosing or treating multiple myeloma
  • Application of RPL8 to preparation of tool for diagnosing or treating multiple myeloma
  • Application of RPL8 to preparation of tool for diagnosing or treating multiple myeloma

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 on the correlation between this gene and multiple myeloma in the prior art as a candidate gene, and considering the results of gene sequencing, select the RPL8 gene (its expression is up-regulated 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 RPL8 gene expression

[0126] 1. siRNA design and synthesis

[0127] siRNA sequence against RPL8:

[0128] siRNA-RPL8:

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

[0130] The antisense strand is 5'-CCUCAAUAAAGUUUGUGUUUA-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 completely to avo...

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Abstract

The invention belongs to the field of biological medicines and discloses application of an RPL8 gene as a diagnosis and treatment marker of multiple myeloma. Experiment proves that the RPL8 gene has significant difference between expression in normal myeloid tissue and expression in multiple myeloma tissue; accordingly, the RPL8 can be used for developing a product for diagnosing multiple myeloma. The invention also discloses application of the RPL8 gene as a treatment target of multiple myeloma. The research achievement provides the theoretical basis for clinicians to make personalized treatment plans, and can provide a novel medicine target for development of medicines for treating 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 by detecting RPL8 abnormality; and a tumor therapeutic agent for inhibiting RPL8 gene or protein. Background technique [0002] Multiple myeloma (Multiple myeloma, MM) is a malignant disease characterized by the accumulation of malignant plasma cells in bone marrow and the appearance of monoclonal protein (M protein) in blood or urine. Clinical features include susceptibility to infection, impaired renal function, anemia, hypercalcemia, and osteolytic lesions. MM accounts for about 10% of all hematological malignancies, and it is more likely to occur in the elderly. With the aging population in my country, the number of MM cases tends to increase. At present, the occurrence of MM is considered to be a progressive process: many patients develop monoclonal glob...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/68G01N33/574A61K45/00A61P35/00
CPCC12Q1/6886A61K45/00C12Q2600/118C12Q2600/158G01N33/57407G01N33/68G01N2333/47
Inventor 杨承刚宋宏涛
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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