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Use of SAV1 (Salvador family WW domain-containing protein 1) gene as hysteromyoma diagnostics and treatment marker

A technology of uterine fibroids and genes, which is applied in the determination/inspection of microorganisms, medical preparations containing active ingredients, biological testing, etc., and can solve the problems of sensitivity and specificity defects of detection methods

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

In the prior art, the methods used to identify the above-mentioned diseases are mostly instrumental examination or according to the doctor's experience, so the detection method has defects in sensitivity and specificity

Method used

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  • Use of SAV1 (Salvador family WW domain-containing protein 1) gene as hysteromyoma diagnostics and treatment marker
  • Use of SAV1 (Salvador family WW domain-containing protein 1) gene as hysteromyoma diagnostics and treatment marker
  • Use of SAV1 (Salvador family WW domain-containing protein 1) gene as hysteromyoma diagnostics and treatment marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Example 1 Differences in the expression of SAV1 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 40 patients with uterine fibroids, and the patients were between 25 and 45 years old.

[0062] 2. Detection of differential expression of the SAV1 gene at the transcriptional level

[0063] 2.1 Extraction of tissue RNA (using NorgenRNA extraction kit)

[0064] 1) In a clean area with less RNase interference, use a mortar containing an appropriate amount of liquid nitrogen to weigh about 20 mg of the isolated tissue sample, and grind it to powder with a pest...

Embodiment 2

[0118] Example 2 SAV1 gene expression plasmid construction

[0119] 1. Construction of SAV1 gene expression vector

[0120] Amplification primers were designed according to the coding sequence of the SAV1 gene (as shown in SEQ ID NO.1). Amplify the coding sequence of the full-length SAV1 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 -SAV1 was used in subsequent experiments.

[0121] 2. Culture and transfection of uterine leiomyoma cells

[0122] After surgical resection of uterine fibroids, 1 cm of each uterine fibroid tissue was immediately taken under aseptic conditions, placed in 10 mL of 3% double-antibody (penicillin, streptomycin) in 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,...

Embodiment 3

[0136] Example 3 Effect of SAV1 gene on proliferation of uterine leiomyoma cells

[0137] MTT assay was used to detect the effect of SAV1 gene on the proliferation ability of uterine leiomyoma cells.

[0138] 1. Steps: trypsinize each group of cells 12 hours after transfection, make a single cell suspension, inoculate 6000 cells per well in a 96-well culture plate, set 7 time points for each group, and set 6 times for each time point. multiple holes. After the cells adhere to the wall, perform the first test: add 20 μl of 5g / L MTT solution to each well, continue to cultivate for 4 hours, aspirate the medium, add 150 μl of DMSO, carefully pipette to fully dissolve the purple-blue precipitate, and use a microplate reader The absorbance value (A value) was measured at a wavelength of 490 nm. Then every 12h detection 1, continuous measurement 72h, a total of 7 times. This experiment was repeated 3 times.

[0139] 2. Statistical methods

[0140] The experiments were all repeat...

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Abstract

The invention discloses a molecular marker, SAV1 (Salvador family WW domain-containing protein 1) gene, for early diagnostics of hysteromyoma and an expression product thereof. By using QPCR (quantitative polymerase chain reaction) and Western blot methods, it is proved that the SAV1 gene has differential expression in hysteromyoma tissues and normal tissues and can be used as an index for the early diagnostics of hysteromyoma. In addition, the invention also discloses the SAV1 gene and the expression product thereof which may be used as targets for treating hysteromyoma and used for guiding the development of new drugs.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the use of SAV1 gene in the diagnosis and treatment of uterine fibroids. Background technique [0002] Uterine fibroids are the most common benign tumors in gynecology. Although most uterine fibroids are asymptomatic and seldom become malignant, they do not need or temporarily do not need treatment, but patients with symptomatic uterine fibroids or those with large Patients should be treated, surgery is the main treatment. Therefore, these patients should be diagnosed accurately before surgery. Because uterine fibroids are easily confused with the following diseases, including pregnancy, uterine hypertrophy, uterine sarcoma, uterine chronic inversion, uterine malformation and pelvic inflammatory mass, it is necessary to differentiate the above diseases. The methods used to identify the above diseases in the prior art are mostly instrumental examination or according to the doctor's ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/68G01N33/574A61K45/00A61P35/00
Inventor 杨承刚边洋杜海威
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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