Application of miR-577 for preparing nephrosis diagnosis marker

A technology of mir-577 and diagnostic reagents, which is applied in the determination/testing of microorganisms, drug combinations, biochemical equipment and methods, etc., and can solve the problems of low early diagnosis rate, lost operation opportunities, hidden onset of renal clear cell carcinoma, etc. question

Inactive Publication Date: 2018-10-09
THE FIRST AFFILIATED HOSPITAL OF ARMY MEDICAL UNIV
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Problems solved by technology

However, clear cell renal cell carcinoma has an insidious onset, most of which have no typical clinical manifestations, and the early diagnosis rate is not high, and about 30% of the patients have metastasized at the time of diagnosis, losing the opportunity for surgery

Method used

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  • Application of miR-577 for preparing nephrosis diagnosis marker
  • Application of miR-577 for preparing nephrosis diagnosis marker
  • Application of miR-577 for preparing nephrosis diagnosis marker

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Embodiment 1 TCGA data analysis

[0047] The mRNA and miRNA data generated by the TCGA database were used. These data were obtained from Primary solid Tumor in the renal clear cell carcinoma case group, and Solid Tissue Normal or Control Analyte* in the control group. The search results are detailed in Table 1 below.

[0048] Table 1

[0049]

[0050] mRNA data analysis results: TCGA database IlluminaHiseq_RNASeqV2 platform has a total of 20531 genes. In this study, 7950 genes that were not easily detected were deleted (only genes without 0 elements in the data were kept). 12581 genes were used for screening of differentially expressed genes. The indicators for screening significantly different mRNAs are: p_value1, |value1-value2|>100, 2361 differentially expressed mRNAs were obtained, 1350 were up-regulated, and 1011 were down-regulated.

[0051] miRNA data analysis results: TCGA database IlluminaHiSeq-miRNASeq platform has a total of 1046 miRNAs. In this stud...

Embodiment 2

[0054] Example 2 Detection of expression of miR-577 and its target genes in clear cell renal cell carcinoma and adjacent tissues by real-time quantitative PCR

[0055] 2.1 Sample acquisition:

[0056] The cancer tissues and paracancerous tissues of 34 patients with clear cell renal cell carcinoma were selected, and all the above specimens were obtained with the consent of the organizational ethics committee.

[0057] 2.2 Extraction of total RNA from samples:

[0058] 1) Take 80 mg tissue block, add 800 μl Lysis / Binding buffer, and use a homogenizer to homogenize the tissue block. During the homogenization process, the samples should be placed on ice to keep the cold state.

[0059] 2) Add 1 / 10 volume of Homogenate Additive to the above homogenized tissue samples, and place on ice for 10 minutes.

[0060] 3) Add an equal volume of water-saturated phenol to the Lysis / Binding buffer, shake for 45 seconds, and centrifuge at 10,000×g for 5 minutes at room temperature.

[0061] ...

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Abstract

The invention relates to application of miR-577 for preparing a nephrosis diagnosis marker, in particular to application of miR-577 and a target gene thereof in preparing a renal clear cell carcinomadiagnosis marker. Database renal clear cell carcinoma mRNA (Ribonucleic Acid) and miRNA data are retrieved, and miR-577 and a target gene regulated and controlled by the miR-577 are selected to carryout molecular verification through data integration, differential expression analysis and targeted analysis. A result indicates that the miR-577 and the target genes CD1D, TMEM133 and SLFN11 regulatedand controlled by the miR-577 can serve as the diagnosis marker of the renal clear cell carcinoma, and therefore good clinic application value is performed.

Description

technical field [0001] The present invention relates to the field of molecular biology, in particular to the application of miR-577 in the preparation of renal disease diagnostic markers, and more specifically to the application of miR-577 in the preparation of renal clear cell carcinoma diagnostic markers. Background technique [0002] miRNAs are a class of single-stranded small RNAs that are reverse complementary to mRNA transcripts of other protein-coding genes, generally 18-25 nt in length. It is produced by DNA transcription, but not translated into protein, but regulates the expression and function of its target genes during protein synthesis. A miRNA is thus a gene that regulates other protein-coding genes. Since miRNA plays an important role in the regulation of gene expression, the difference in miRNA expression can cause a variety of human diseases. The currently known disease systems related to miRNA expression disorders include: tumors, rheumatic diseases, metab...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886A61K45/00A61P35/00
CPCA61K45/00A61P35/00C12Q1/6886C12Q2600/158C12Q2600/178
Inventor 申兵冰周强梅玫于艳杰刘宏赵洪雯
Owner THE FIRST AFFILIATED HOSPITAL OF ARMY MEDICAL UNIV
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