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

Application of gene SUCLG1 or expression products thereof and detection kit

A technology for detection kits and expression products, applied in the biological field, can solve the problems of unclear specific mechanism of renal cell carcinoma

Pending Publication Date: 2021-09-03
深圳市龙华区人民医院
View PDF6 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the specific mechanism of the occurrence, development and metastasis of renal cell carcinoma is still unclear

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 gene SUCLG1 or expression products thereof and detection kit
  • Application of gene SUCLG1 or expression products thereof and detection kit
  • Application of gene SUCLG1 or expression products thereof and detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Expression of SUCLG1 in Different Renal Cancer Cell Lines

[0033] Place the cell culture dish on ice, remove the medium, and wash the cells three times with PBS. use Kit (Fast gene, Inc.; Cat. No. 220010) was used to extract total RNA from OSRC-2, A498, 786-D, Caki-1, ACHN, and 769-P cells, respectively. Following the manufacturer's instructions, use a DNA eraser with The RT kit (Takara BioInc.) reverse-transcribed the extracted RNA into cDNA, respectively using cDNA as a template and using SEQ ID NO: 1 and SEQ ID NO: 2 as primers, and performing real-time fluorescent quantitative PCR according to conventional methods and steps. see results figure 1 .

[0034] forward 5'-GAGCAACGGCTTCTGTCATTT-3' (SEQ ID NO: 1);

[0035] reverse 5'-TGCTTGACTCGTACCATGTCC-3' (SEQ ID NO: 2).

[0036] According to the above steps, the total RNA of kidney cancer tissue was extracted, the reverse transcribed cDNA was used as a template, and SEQ ID NO: 1 and SEQ ID NO: 2 were...

Embodiment 2

[0039] Example 2 Effect of overexpression of SUCLG1 on the proliferation of renal cancer cells

[0040] In order to measure cell viability, the plasmid pSUCLG1 overexpressing SUCLG1 was constructed to transfect OSRC-2 and A498 cell lines, respectively, and the transfected cells were cultured and counted in a 96-well plate (1000 cells per well). The time for cell culture and counting is 0, 24, 48, 72, and 96 hours, and the cell viability is measured using the cell counting kit CCK8 analysis kit, and the operation is performed according to the steps of the kit instructions. see results Figure 4 .

[0041] Use the edU cell proliferation detection kit to detect the cell proliferation of the cells cultured for 24 hours, and perform the test according to the steps in the instructions, and the results are shown in Figure 5 , where, vector is the control group transfected with empty plasmid.

[0042] Depend on Figure 4~5 The results showed that the overexpression of SUCLG1 gene...

Embodiment 3

[0043] Example 3 Effect of Overexpression of SUCLG1 on Migration and Invasion of Renal Cancer Cells

[0044] (1) transwell experiment

[0045] Construct OSRC-2 and A498 cells overexpressing SUCLG1, digest with trypsin, centrifuge at 900×g for 3 minutes at room temperature, discard the supernatant, resuspend the sample in RPMI-1640 medium, and incubate at 900×g Centrifuge for 3 minutes and wash cells with PBS. The cells were resuspended in 500 μl RPMI-1640 medium, and the cell density was measured with a hemocytometer. In the invasion assay, the Transwell membrane was wrapped with Matrigel. A total of 2×10 4 cells were placed in the upper chamber of the 8 μm microwell Transwell insert, 500 μl of RPMI-1640 supplemented with 10% FBS was added in the lower chamber, and then the cells were cultured. Migration and invasion were determined by counting the number of cells that successfully migrated through the cell membrane (migration) or through the Matrigel matrix (invasion). Af...

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 the technical field of biology and particularly relates to application of a gene SUCLG1 as a tumor marker and a detection reagent. The invention provides application of the gene SUCLG1 in preparation of a renal carcinoma detection reagent as a marker. According to the application, discovered by researches, the proliferation of renal carcinoma cells and the migration and invasion of the renal carcinoma cells can be effectively inhibited through over-expressing the gene SUCLG1; and through detecting an expression level of the gene SUCLG1 in a sample, whether an individual suffers from a renal carcinoma or not can be accurately judged or the risk of suffering from the renal carcinoma can be accurately evaluated, and a relationship between the expression level of the gene SUCLG1 and renal carcinoma onset is discovered and verified for the first time.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an application detection kit for SUCLG1 gene or its expression product. Background technique [0002] Kidney cancer is one of the most common urological tumors in the world. Kidney cancer is on the rise year by year worldwide, with an annual incidence of 2%. Statistics show that the incidence of kidney cancer in developed countries is higher than that in backward countries. The incidence of kidney cancer in men is about 1.58 times that in women. The incidence of RCC in my country is relatively low, but the incidence of RCC has been rising rapidly in recent years. In 2019, there were an estimated 73,820 new cases and 14,770 deaths from kidney cancer in the United States. The morbidity and mortality of RCC in my country are also on the rise. In 2015, there were an estimated 66,800 new cases and 23,400 deaths. According to the prediction of the World Health Organization, the progn...

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): C12Q1/6886C12Q1/686G01N33/574G01N33/68C12N15/11
CPCC12Q1/6886C12Q1/686G01N33/57438G01N33/68C12Q2600/158C12Q2561/113C12Q2563/107C12Q2545/114
Inventor 陈业达陈应坚梁碧玉何伟明王杰燕
Owner 深圳市龙华区人民医院
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