NUDT11 genes and expression products thereof serving as diagnosis targets of UL (uterine leiomyoma)

A technology of uterine fibroids and gene expression, which can be used in gene therapy, microbial determination/examination, biological testing, etc., and can solve problems such as unclear causes and mechanisms

Active Publication Date: 2016-06-08
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, its etiology and mechanism are still unclear

Method used

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  • NUDT11 genes and expression products thereof serving as diagnosis targets of UL (uterine leiomyoma)
  • NUDT11 genes and expression products thereof serving as diagnosis targets of UL (uterine leiomyoma)
  • NUDT11 genes and expression products thereof serving as diagnosis targets of UL (uterine leiomyoma)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Differential expression of embodiment 1 NUDT11 gene

[0061] 1. Experimental materials:

[0062] 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.

[0063] 2. Acquisition of RNA from uterine fibroid tissue and normal muscle epithelial tissue

[0064] Total RNA was extracted from uterine fibroid tissue and normal myometrial tissue using Trizol one-step method, and the purity of the RNA solution was determined by reading the absorbance values ​​at 260nm and 280nm (A) by NanodropND-1000. The integrity of RNA was detected by electrophoresis on 1% formaldehyde denatured agarose ge...

Embodiment 2

[0073] Differential expression of embodiment 2 NUDT11 protein

[0074] 1. The research object is the same as in Example 1.

[0075] 2. Extract tissue total protein

[0076] Follow the instructions of the EpiQuik Tissue / Cell Total Protein Extraction Kit for protein extraction.

[0077] 3. Western blot detection

[0078] The extracted protein was quantitatively subjected to SDS-PAGE electrophoresis, followed by membrane transfer, blocking, primary antibody incubation, secondary antibody incubation, and color development.

[0079] 4. Statistical processing

[0080] The gray value of the protein band was analyzed using ImageJ software, and β-actin was used as an internal reference to normalize the gray value of the NUDT11 protein band. The results and data are expressed in the form of mean ± standard deviation, and SPSS13.0 statistical software is used for statistical analysis. The difference between the two is tested by t test, and it is considered statistically significant w...

Embodiment 3

[0083] Embodiment 3 inhibits NUDT11 gene expression

[0084] 1. siRNA design and synthesis

[0085]siRNA sequence against NUDT11:

[0086] siRNA1-NUDT11:

[0087] The sense strand is 5'-AAUGCUAACCGAAUCUUCCCA-3' (SEQ ID NO.7);

[0088] The antisense strand is 5'-GGAAGAUUCGGUUAGCAUUGG-3' (SEQ ID NO.8),

[0089] siRNA2-NUDT11:

[0090] The sense strand is 5'-ACAAUACUGAACAUCUUUGCU-3' (SEQ ID NO.9);

[0091] The antisense strand is 5'-CAAAGAUGUUCAGUAUUGUGC-3' (SEQ ID NO.10),

[0092] siRNA3-NUDT11:

[0093] The sense strand is 5'-UCUACUAGCACCAGAUUGGAA-3' (SEQ ID NO.11);

[0094] The antisense strand is 5'-CCAAUCUGGUGCUAGUAGAGU-3' (SEQ ID NO.12)

[0095] Negative control siRNA sequence (siRNA-NC):

[0096] The sense strand is 5'-CGUACGCGGAAUACUUCGA-3' (SEQ ID NO.13);

[0097] The antisense strand is 5'-UCGAAGUAUUCCGCGUACG-3' (SEQ ID NO.14).

[0098] 2. Culture and transfection of uterine leiomyoma cells

[0099] After surgical resection of uterine fibroids, 1 cm of each ...

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Abstract

The invention discloses NUDT11 genes and expression products thereof serving as molecular markers for diagnosis of UL (uterine leiomyoma). Whether a subject suffers from UL can be judged and whether the subject has the risk of suffering from UL can be diagnosed by detecting the content of the NUDT11 genes and the expression products thereof in tissue. Researches on proliferation conditions of UL cells cultured in vitro show that expression inhibition of the NUDT11 genes can inhibit UL cell proliferation, proliferation of the UL cells can be inhibited by inhibiting the function of NUDT11 protein, and research results show that the NUDT11 genes and the expression products thereof are potential medicine targets for treatment of UL.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the use of NUDT11 gene in the diagnosis and treatment of uterine fibroids. Background technique [0002] Uterine leiomyoma (uterine leiomyoma, UL) is the most common benign tumor in the female reproductive system. It is more common in women aged 30 to 50, and the incidence rate among women aged 40 to 50 is as high as 51.2% to 60.0%. However, its etiology and mechanism are still unclear. With the study of histogenesis, endocrinology and molecular genetics of uterine fibroids, the pathogenesis of uterine fibroids and the factors that promote the growth of uterine fibroids have been further understood. In recent years, the study of susceptibility genes has gradually become a hot spot in the study of uterine fibroids. Determining the susceptibility genes of uterine fibroids to determine the susceptible population will lay the foundation for the prevention and treatment of diseases, an...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/68G01N33/574A61K45/00A61K48/00A61K31/713A61K39/395A61P35/00
CPCA61K31/713A61K39/395A61K45/00C12Q1/6886C12Q2600/158G01N33/57411G01N33/68
Inventor 杨承刚边洋杜海威
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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