Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Application of SPAG6 gene serving as diagnosis and treatment marker for tumor ovarii

A technology of ovarian tumors and genes, applied in gene therapy, antineoplastic drugs, genetic engineering, etc.

Active Publication Date: 2017-03-29
WUHAN UNIV OF SCI & TECH
View PDF4 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, microtubule-binding drugs, such as paclitaxel, have played an important role in the treatment of ovarian cancer. However, due to drug tolerance and side effects, such as allergic reactions, bone marrow suppression, cardiovascular and cerebrovascular system lesions, nausea, vomiting, joint Pain and hair loss, among other things, limit these drugs from being more effective

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Application of SPAG6 gene serving as diagnosis and treatment marker for tumor ovarii
  • Application of SPAG6 gene serving as diagnosis and treatment marker for tumor ovarii
  • Application of SPAG6 gene serving as diagnosis and treatment marker for tumor ovarii

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Example 1 Differences in SPAG6 gene expression between normal tissue and ovarian tumor tissue.

[0065] 1. Materials and methods.

[0066] 1.1 The source of the patient.

[0067] Tumor and paracancerous normal tissue samples from patients undergoing ovarian cancer surgery were collected from several hospitals in Wuhan. All patients signed informed consent, which was approved by the Medical Ethics Committee of Wuhan University of Science and Technology.

[0068] 2. Pyrosequencing method to detect methylation of SPAG6 promoter CpG island.

[0069] 2.1 Materials.

[0070] 2.1.1 Reagents and kits.

[0071]

[0072] 2.1.2 Instruments.

[0073]

[0074] 2.2 Method.

[0075] 2.2.1 Methylation modification

[0076] (1) Use sulfite reagent to dissolve DNA, add 800 μl RNase-free water to each sulfite mixture, and mix well;

[0077] (2) Prepare the following reagents according to Table 1 in a 200 μl PCR thin-walled tube;

[0078] Table1. sulfite modification reactio...

Embodiment 2

[0134] Example 2 Differences in SPAG6 gene expression between normal ovarian epithelial cells and ovarian cancer epithelial cells.

[0135] 1. Detect the differential expression of SPAG6 gene at the transcriptional level.

[0136] 1.1 Total RNA extraction

[0137] (1) Preparation: Trizol, chloroform (chloroform), isopropanol, 75% ethanol containing DEPC (1‰), PBS buffer solution, EP tube 1.5ml and 200ul are treated with double distilled water containing 1‰DEPC After high-pressure drying, 1ml, 200ul, and 10ul of the pipette tip were treated with DEPC water and then high-pressure dried. The DEPC water was prepared at a ratio of 1:1000 and placed for more than 4 hours;

[0138] (2) Operation steps: Discard the culture medium, add 1ml TRIzol, and use a homogenizer for homogenization. The sample volume should not exceed the volume of TRIzol by 10℅. Place the homogenized sample at room temperature (15-30° C.) for 5 minutes to completely separate the nucleic acid-protein complexes...

Embodiment 3

[0160] Example 3 To study the effect of SPAG6 gene expression inhibition on the malignant biological traits of tumor cells.

[0161] The steps of transfecting CHO cells / COS-7 cells with recombinant plasmid GFP-Spag6L / pcDNA3-Spag6L are as follows:

[0162] (1) 24 hours in advance, inoculate CHO cells / COS-7 cells into six-well plates (six-well plates used for immunofluorescence experiments need to put cell slides in each well in advance), and wait until the cell growth density is 50%- 70% (1×10 5 individual / ml) transfection;

[0163] (2) Two hours before transfection, replace the cells with fresh cell culture medium;

[0164] (3) According to the concentration of plasmid DNA, take 0.8 μg plasmid DNA (about 8-10 μl) and dilute it with 25 μl DMEM culture medium, mix well, and make dilution A;

[0165] (4) Dilute 2 μl of transfection reagent with 25 μl of cell culture medium, mix well to make dilution B, and let it stand at room temperature for 5 minutes;

[0166] (5) Add all t...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to application of an SPAG6 gene serving as a diagnosis and treatment marker for tumor ovarii. The QPCR and Western blot methods are utilized for proving that the SPAG6 gene expresses differently in tumor ovarii tissue and normal tissue, in vitro growth curves and in vitro taxol growth inhibition tests are utilized for detecting influences of the SPAG6 gene on reproductive development of an ovarian carcinoma cell SKOV-3, and the SPAG6 can serve as an indicator for early diagnosis of the tumor ovarii. According to application of the SPAG6 gene serving as the diagnosis and treatment marker for tumor ovarii, through SPAG6 tests on ovarian epidermal tumors, new target spots are provided for diagnosis and treatment of the tumor ovarii and the like, research and development of new drugs are guided, and the curative ratio and survival rate of tumor patients are increased accordingly.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to the application of SPAG6 gene in the diagnosis and treatment of ovarian tumors. Background technique [0002] Microtubules are a fibrous structure ubiquitous in eukaryotic cells, formed by the polymerization of tubulin and a small amount of microtubule-binding proteins, and are one of the main components of the cytoskeleton. Microtubules play a very important role in the body, mainly involved in cell movement, organelle positioning, intracellular material transport, eukaryotic cell mitosis process and cell signal transmission. [0003] In the cell, microtubules exist in two forms: cytoplasmic microtubules and spindle microtubules. These two types of microtubules are in dynamic equilibrium of assembly and disassembly in different cell cycles. Any factors that make this balance unbalanced Can destroy the microtubule structure, so as to affect cell function. Tumor cells have t...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12Q1/68G01N33/68G01N33/574A61K48/00A61K31/7088A61P35/00
CPCA61K31/7088C12N15/113C12N2310/14C12Q1/6886C12Q2600/154C12Q2600/158G01N33/57449G01N33/68C12N2310/531
Inventor 张志兵石玉琴张玲柳赟昊付国庆江高峰周婷李玉红
Owner WUHAN UNIV OF SCI & TECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com