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Application of miRNA-424 as diagnostic marker of pituitary adenoma

A technology of miRNA-424 and diagnostic markers, which is applied in the field of biomedicine and can solve problems such as deficiency

Inactive Publication Date: 2021-03-16
YANGTZE DELTA REGION INST OF TSINGHUA UNIV ZHEJIANG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Large-scale studies using miRNA expression signatures to differentiate pituitary tumors are still lacking

Method used

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  • Application of miRNA-424 as diagnostic marker of pituitary adenoma
  • Application of miRNA-424 as diagnostic marker of pituitary adenoma
  • Application of miRNA-424 as diagnostic marker of pituitary adenoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 qPCR verification of differentially expressed miRNA-424

[0047] 1. Sample acquisition

[0048] 5 cases of normal pituitary gland tissues and 53 cases of pituitary tumor tissues including all 5 subtypes were collected. All the above specimens were obtained with the consent of the organizational ethics committee.

[0049] 2. Extraction of total RNA from samples

[0050] Total RNA in advance using QIAGEN Tissue RNA Extraction Kit.

[0051] Specific steps are as follows:

[0052] 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 tissue sample, and grind it to powder with a pestle;

[0053] 2) Transfer the sample to a RNase-free 2mL centrifuge tube;

[0054] 3) Add 300 μL Lysis solution, place in a homogenizer, and grind thoroughly for 1-5 minutes;

[0055] 4) 12000g, 4°C, centrifuge for 10min, transfer the supernatant to a new 1.5mL centrifuge tube;

[0056] 5) Add ...

Embodiment 2

[0087] Example 2 Identification of miRNAs specifically expressed in pituitary tumors and their key target genes

[0088] 1. Bioinformatics method to predict the potential key target genes of miRNA-424

[0089]Combine multiple methods to improve the accuracy of analysis, use TargetScan, miRecords and PicTar and other software for prediction, and take the intersection of several prediction software results for analysis. Because this bioinformatics analysis may obtain a large number of possible target genes, it is necessary to focus on selecting potential target genes that may function in the occurrence and development of pituitary tumors for analysis on the basis of existing pituitary tumor research. For one miRNA, multiple possible target genes can be analyzed and verified simultaneously to increase the success rate.

[0090] 2. Results

[0091] The possible key target genes of miR-424 were predicted by bioinformatics analysis. The results showed that the 3' non-coding regio...

Embodiment 3

[0092] Example 3 Detection of expression of miRNA-424 and HMGA1 in pituitary tumor cells by qPCR and detection of HMGA1 protein expression by western blotting

[0093] 1. Cell culture

[0094] GT1-1 cells were routinely cultured in F-12 medium containing 5% fetal bovine serum and 5% horse serum at 37°C, 5% CO 2 and cultured under saturated humidity conditions.

[0095] 2. Cell transfection

[0096] The GT1-1 cells were divided into four groups, the experimental group was transfected with miRNA-424 mimics and miRNA-424 inhibitors, the negative control group was transfected with random control sequences, and the liposome control group was transfected with LipofectamineTM 3000 lipid In vivo transfection reagent group, and the blank control group was the non-transfection group. The transfection reagent LipofectamineTM 3000 was used for transfection, and the transfection method was referred to the instructions. The working concentration of both the control group and the experim...

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Abstract

The invention discloses an application of miRNA-424 as a diagnostic marker of pituitary adenoma and preparation or screening of drugs for treating pituitary adenoma. The invention discovers for the first time that the expression level of miRNA-424 is related to the occurrence of pituitary adenoma. By detecting the expression level of miRNA-424 in a patient, whether the subject has the risk of pituitary adenoma can be judged, thereby guiding the clinician to provide a prevention scheme or a treatment scheme for the subject.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to miRNA diagnostic markers for pituitary tumors, and in particular to the application of miRNA-424 as a diagnostic marker for pituitary tumors. Background technique [0002] MicroRNAs (miRNAs) are relatively recently discovered non-coding RNAs with regulatory functions in eukaryotes. Mature miRNA is a single-stranded RNA molecule containing approximately 19-23 nucleotides, which is produced by a series of enzymatic cleavage processes from a long primary transcript. [0003] miRNAs are mainly involved in the regulation of gene expression at the post-transcriptional level. miRNA binds to the miRNA recognition element in the 3' untranslated region of the target gene through sequence complementarity, preventing the translation of the target gene by blocking the protein translation machinery or by separating the mRNA transcript from the ribosome. miRNAs can also induce the degrada...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886A61K31/7105A61P35/00
CPCA61K31/7105A61P35/00C12Q1/6886C12Q2600/118C12Q2600/136C12Q2600/158C12Q2600/178
Inventor 李旭辉徐文飞柴颖杰王星华
Owner YANGTZE DELTA REGION INST OF TSINGHUA UNIV ZHEJIANG
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