Diagnosis and treatment markers for hysteromyoma

A technology for uterine fibroids and products, which can be applied in gene therapy, biological testing, anti-tumor drugs, etc., and can solve the problems of early diagnosis of uterine fibroids

Active Publication Date: 2016-04-20
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0013] However, the above detection methods canno

Method used

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  • Diagnosis and treatment markers for hysteromyoma
  • Diagnosis and treatment markers for hysteromyoma
  • Diagnosis and treatment markers for hysteromyoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Differential expression of embodiment 1GPSM2 gene

[0073] 1. Experimental materials:

[0074] 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 40 patients with uterine fibroids, and the patients were between 25 and 45 years old.

[0075] 2. Gene chip screening

[0076] Relying on the technical service of the gene chip analysis platform provided by Biobio, the AFFX human genome expression profile chip (AFFXHumanGenomeU133Plus2.0Array, Affimetrix Company) was selected to complete the gene chip screening analysis of tissue RNA samples, its working principle and the entire operation process Referring to the company's conventional gene chip analysis method, the m...

Embodiment 2

[0085] Differential expression of embodiment 2GPSM2 protein

[0086] 1. The research object is the same as in Example 1.

[0087] 2. Extraction of total tissue protein

[0088] Follow the instructions of the EpiQuik Tissue / Cell Total Protein Extraction Kit for protein extraction.

[0089] 3. Western blot detection

[0090] The total protein was quantified by the Brandford method, take an appropriate amount and mix it with the sample buffer, boil for 5 minutes, and cool for 5 minutes; take 30pg of protein and load it on a prepared 15% polyacrylamide gel for electrophoresis, start with a constant voltage of 80V, and see the Marker Increase to 120V; take out the gel after electrophoresis, use Bio.Rad semi-dry transfer system to transfer at 100V for 50min; after the transfer is completed, wash once with 1xPBS, immerse in blocking solution, 40C overnight; pour off blocking solution, add Western wash Wash with the membrane washing solution for 5-10 minutes, add the primary antibo...

Embodiment 3

[0095] Embodiment 3 suppresses GPSM2 gene expression

[0096] 1. siRNA design and synthesis

[0097] siRNA sequence against GPSM2:

[0098] siRNA1-GPSM2:

[0099] The sense strand is 5'-UCACUAAUGAUAGGUUUUCCU-3' (SEQ ID NO.7);

[0100] The antisense strand is 5'-GAAAACCUAUCAUUAGUGACU-3' (SEQ ID NO.8),

[0101] siRNA2-GPSM2:

[0102] The sense strand is 5'-AGUAAAGUAUAUGUAUUUCCA-3' (SEQ ID NO.9);

[0103] The antisense strand is 5'-GAAAUACAUAUACUUUACUUC-3' (SEQ ID NO.10),

[0104] siRNA3-GPSM2:

[0105] The sense strand is 5'-AUUAUGGAGUUAUUUUGUGCUG-3' (SEQ ID NO.11);

[0106] The antisense strand is 5'-GCACAAAUAACUCCAUAAUGU-3' (SEQ ID NO.12)

[0107] Negative control siRNA sequence (siRNA-NC):

[0108] The sense strand is 5'-CGUACGCGGAAUACUUCGA-3' (SEQ ID NO.13);

[0109] The antisense strand is 5'-UCGAAGUAUUCCGCGUACG-3' (SEQ ID NO.14).

[0110] 2. Culture and transfection of uterine leiomyoma cells

[0111] After surgical resection of uterine fibroids, 1 cm of each u...

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Abstract

The invention discloses GPSM2 genes which can be used as the molecular markers of the early diagnosis of hysteromyoma. Experiments prove that the GPSM2 gene expression in hysteromyoma tissue is increased evidently as compared with that of normal muscular layer tissue. RNA interference experiments prove that the GPSM2 can affect the proliferation of hysteromyoma cells. Medicine capable of inhibiting the GPSM2 gene expression can be developed, and clinical prevention and treatment of the hysteromyoma can be achieved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of GPSM2 gene in the diagnosis and treatment of hysteromyoma. Background technique [0002] Uterine fibroids are the most common benign tumors in female reproductive organs and one of the most common tumors in the human body, also known as fibromyomas and uterine fibroids. Since uterine fibroids are mainly formed by the proliferation of uterine smooth muscle cells, a small amount of fibrous connective tissue exists as a supporting tissue, so it is more accurate to call it uterine leiomyoma, or uterine fibroids for short. Uterine fibroid inspection methods commonly used in the prior art include: [0003] 1. Ultrasonography [0004] It is the most commonly used auxiliary diagnostic method at present. It can show the enlarged uterus, irregular shape, the number, location, size of fibroids, and whether the interior of fibroids is uniform or liquefied, cystic, etc. Ult...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/68G01N33/574A61K45/00A61K48/00A61K39/395A61K31/7088A61P35/00
CPCA61K31/7088A61K45/00A61K2039/505C12Q1/6886C12Q2600/158G01N33/57442G01N33/68
Inventor 杨承刚孙耀兰石小峰
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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