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New uses of mir-3620

A technology of colon adenocarcinoma and preparation, applied in the new application field of mir-3620 and its mature miRNA in the diagnosis and treatment of colon adenocarcinoma, which can solve the problems affecting the normal physiological function of cells and diseases

Active Publication Date: 2019-07-12
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The abnormality of miRNA expression and activity is very likely to affect the normal physiological function of cells and lead to the occurrence of diseases

Method used

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  • New uses of mir-3620
  • New uses of mir-3620
  • New uses of mir-3620

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] The collection of embodiment 1 sample and total RNA extraction

[0053] From January 2014 to August 2015, 6 cases of cancer tissues and corresponding paracancerous tissues were collected from patients with colon adenocarcinoma. The patients did not receive radiotherapy and chemotherapy before the operation, and the materials were collected during the operation, stored in liquid nitrogen immediately, and then transferred to -80°C for long-term storage for RNA extraction. The specimen was diagnosed as colon cancer by pathological diagnosis.

[0054] RNA extraction standards: RNA purity: OD260 / 280≧1.8, 28S / 18S≧1; RNA integrity: RIN value≧7.0. RNA integrity detection method: Agilent 2100 (RNA 6000Nano kit), agarose gel electrophoresis (agarose gel concentration: 1% agarose gel; voltage: 5V / cm; time: 20min).

Embodiment 2

[0055] Example 2 Sequencing and Data Integration Analysis

[0056] Sequencing: The miRNA was sequenced using llumina Hiseq2500 / Miseq second-generation high-throughput sequencing technology, and the data processing was completed through the processes of removing adapters, removing low-quality, and decontaminating to obtain the final data.

[0057] Meta-analysis: The nationalwidechildrens.org_clinical_patient_coad file of the TCGA database recorded the clinical information of 459 patients with colon adenocarcinoma. This project excludes 62 patients whose history_other_malignancy or history_neoadjuvant_treatment is Yes, and 337 patients whose histologic_diagnosis is ColonAdenocarcinoma are included in the study (60 patients whose histologic_diagnosis is Colon MucinousAdenocarcinoma, [Not Available], [Discrepancy] are not included in the study). The miRNA data in the TCGA database of 337 patients with colon adenocarcinoma (COAD) were used for analysis. These data are from the Pri...

Embodiment 3

[0059] Example 3 Detection of miR-3620-3p expression in colon adenocarcinoma tissue by Real-time PCR

[0060] 1 sample collection:

[0061] Cancer tissues and paracancerous tissues of 36 patients with colon adenocarcinoma (collection time from January 2014 to August 2015).

[0062] 2 total RNA extraction:

[0063] Removal of RNase for related experimental items:

[0064] ① Rinse and soak all glassware with DEPC before use, 120°C for 20 minutes under high pressure, and 180°C for more than 2 hours.

[0065] ②Plastic utensils (such as: EP tubes / tips) need to be soaked in 0.1% DEPC water overnight before use, and then the liquid is controlled to dry, 120°C and high pressure for 20 minutes, and dried in an oven for later use.

[0066] leukapheresis

[0067] (1) Take 2m1 anticoagulated peripheral blood (blood collection time does not exceed 3h);

[0068] (2) Add an equal volume of sterile PBS to the peripheral blood and mix thoroughly to form a cell suspension;

[0069] (3) Add ...

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Abstract

The invention relates to a novel application of mir-3620, in particular to novel applications of the mir-3620 and mature miRNA thereof in diagnosing and treating colon adenocarcinoma. According to the novel application of the mir-3620, next generation sequencing is conducted on colon adenocarcinoma tissues and para-carcinoma tissues, so that data is obtained, and meanwhile, the obtained data is integrated and analyze in combination with existing data of a database, so that miRNA expression data is obtained and candidate mir-3620 is screened out. Further clinical verification and cell experiment results indicate that miR-3620-3p achieves low expression in the cancer tissues, and the over-expression of the miR-3620-3p can inhibit in vitro proliferation capacity of the colon adenocarcinoma cells. The invention provides a novel colon adenocarcinoma diagnosis and treatment gene for clinical application and lays a foundation for further research and development of related reagents.

Description

technical field [0001] The present invention relates to the field of tumor molecular diagnosis, in particular to the new application of mir-3620 and its mature miRNA, and more specifically to the new application of mir-3620 and its mature miRNA in the diagnosis and treatment of colon adenocarcinoma. Background technique [0002] miRNA is an endogenous non-coding small molecule that regulates gene expression by specifically binding to target genes. A single miRNA can target hundreds of target genes, and the interaction between a large number of miRNAs and target genes plays a crucial role in the regulation of cancer-related signaling pathways. miRNAs function as regulatory feedback and buffers in normal cell physiology, serving as key factors for fine-tuning target gene protein levels and preventing them from exceeding the normal physiological range. In addition, miRNA is also a switch between different molecular signaling pathways. The abnormality of miRNA expression and a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886C12N15/113A61K45/00A61K31/713A61P35/00
CPCA61K31/713A61K45/00C12Q1/6886C12Q2600/178
Inventor 杨承刚董东杜海威
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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