Molecular marker for diagnosing and treating hysteromyoma

A uterine fibroid, a pair of technology, applied in gene therapy, analytical materials, biological testing, etc., can solve the problems of patients feeling uncomfortable and poor sensitivity

Active Publication Date: 2016-05-25
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the above-mentioned physical methods have the disadvantages of poor sensitivity or making patients feel uncomfortable, so the emergence of new diagnostic methods is expected

Method used

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  • Molecular marker for diagnosing and treating hysteromyoma
  • Molecular marker for diagnosing and treating hysteromyoma
  • Molecular marker for diagnosing and treating hysteromyoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Example 1 Differences in the expression of SYBU gene in normal muscle layer tissue and uterine leiomyoma tissue

[0060] 1. Experimental materials:

[0061] Uterine fibroid tissue and adjacent normal muscle layer tissue were aseptically collected from patients undergoing total hysterectomy for uterine fibroids. The patient had not received hormone therapy within 3 months before operation, and all patients were pathologically diagnosed as uterine fibroids after operation. , the experimental materials were taken from 5 patients with uterine fibroids, and the patients were between 25 and 45 years old.

[0062] 2. Detection of differential expression of SYBU gene

[0063] 2.1 Extraction of tissue RNA

[0064] Use QIAGEN's RNApreppureTissueKit (Animal Tissue Total RNA Extraction Kit (DP431)) to extract according to the operating instructions.

[0065] 2.2 Quality analysis of RNA samples (NanoDrop1000 spectrophotometer)

[0066] NanoDrop1000 spectrophotometer detects RNA ...

Embodiment 2

[0078] Example 2 Verification of Differentially Expressed Genes

[0079] 1. Experimental materials:

[0080] Uterine fibroid tissue and adjacent normal muscle layer tissue were aseptically collected from patients undergoing total hysterectomy for uterine fibroids. The patient had not received hormone therapy within 3 months before operation, and all patients were pathologically diagnosed as uterine fibroids after operation. , the experimental materials were taken from 50 patients with uterine fibroids, and the patients were between 25 and 45 years old.

[0081] 2. Detection of differential expression of SYBU gene

[0082] 2.1 Extraction of tissue RNA

[0083] Use QIAGEN's RNApreppureTissueKit (Animal Tissue Total RNA Extraction Kit (DP431)) to extract according to the operating instructions.

[0084] 2.2 Reverse transcription

[0085]1 μg of total RNA was reverse-transcribed to synthesize cDNA using reverse transcription buffer. Using a 25 μl reaction system, take 1 μg of...

Embodiment 3

[0108] Example 3 SYBU gene expression plasmid construction

[0109] 1. Construction of SYBU gene expression vector

[0110] Amplification primers were designed according to the coding sequence of the SYBU gene (as shown in SEQ ID NO.1). Amplify the coding sequence of the full-length SYBU 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 -SYBU was used in subsequent experiments.

[0111] 2. Culture and transfection of uterine leiomyoma cells

[0112] After surgical resection of uterine fibroids, 1 cm of uterine fibroid tissue was immediately taken under aseptic conditions, placed in 10 mL of 3% double-antibody (penicillin, streptomycin) PBS, and sent to the cell culture room as soon as possible in an airtight ice bath. Uterine leiomyoma tissue was rinsed with 3% double-antibody PBS solution, and tri...

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Abstract

The invention discloses a molecular marker for diagnosing and treating hysteromyoma. QPCR and Western blot methods prove remarkable difference of expressions of SYBU genes in the hysteromyoma tissue and normal tissues. Moreover, in-vitro cell culture experiments of the invention prove that the expression of the SYBU genes can inhibit the proliferation of hysteromyoma cells while promoting apoptosis, so that the SYBU genes and expression products thereof can be used for preparing a medicine for treating hysteromyoma and are widely and clinically applied.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of SYBU gene in the diagnosis and treatment of uterine fibroids. Background technique [0002] Uterine fibroids (alias: uterine benign tumor English: myomaofuterus), is the most common benign tumor of female genitalia. Most are asymptomatic, and a few manifest as vaginal bleeding, palpable abdominal masses, and compression symptoms. Multiple uterine fibroids are common. The exact cause of this disease is unknown. Modern western medicine adopts sex hormones or surgery, and there is no other ideal therapy. , Due to the obvious decline of ovarian function, most of the fibroids shrink by themselves. [0003] The diagnosis of uterine fibroids can be initially judged by some symptoms of uterine fibroids, and then further examinations can be made, but the diagnosis cannot be made by gynecological examination alone. First of all, patients with uterine fibroids will feel ab...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/68G01N33/574A61K45/00A61K48/00A61K38/17A61P35/00A61P15/00
CPCA61K38/1709A61K45/00A61K48/005C12Q1/6886C12Q2600/158G01N33/57442G01N33/68
Inventor 杨承刚宋宏涛李曙光
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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