Application of miR-3613 as miRNA marker in distinguishing metastasis and non-metastasis of lung squamous carcinoma

A lung squamous cell carcinoma, tiny technology, used in the field of biomedicine

Active Publication Date: 2016-05-25
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The lack of effective early diagnosis and treatment methods for non-small cell lung cancer metastasis and drug resistance is the bottleneck in the current prevention and treatment

Method used

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  • Application of miR-3613 as miRNA marker in distinguishing metastasis and non-metastasis of lung squamous carcinoma
  • Application of miR-3613 as miRNA marker in distinguishing metastasis and non-metastasis of lung squamous carcinoma
  • Application of miR-3613 as miRNA marker in distinguishing metastasis and non-metastasis of lung squamous carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Example 1 Solexa sequencing screening for miRNAs associated with lung squamous cell carcinoma

[0055] 1. Sample collection

[0056] Collect 40 lung squamous cell carcinoma tissues (including 20 samples with metastasis and 20 samples without metastasis). The above samples are surgical resection specimens of patients with lung squamous cell carcinoma. All the above specimens were obtained with the consent of the organizational ethics committee. The clinical data of the tissue samples include: gender, age, tumor size, pathological grade (Edmonson), metastasis or recurrence, etc.

[0057] 2. RNA extraction: dry-bake the pestle and homogenizer at 200°C for 4 hours to remove RNase and cool down; add liquid nitrogen to pre-cool, quickly remove the tissue from the liquid nitrogen, and grind it into powder; use a spatula Put the tissue into a homogenizer pre-added with TRIzol reagent, and homogenize for a few minutes; transfer the homogenized liquid into an RNase-free centrif...

Embodiment 2

[0062] Example 2 Expression of miR-3613 in lung squamous cell carcinoma cell lines

[0063] 1. Cell culture

[0064] The lung squamous cell line NCI-H520, NCI-H596, NCI-H2170, NCI-H226 were cultured in DMEM medium and 10% fetal bovine serum, and the human lung epithelial cell line BEAS-2B was cultured in BEGM medium and 10% Cultured in fetal bovine serum at 37°C, 5% CO 2 in the incubator.

[0065] 2. QPCR

[0066] 2.1 Extraction of total cellular RNA: Extraction of total cellular RNA was performed using the RNA extraction kit from QINGEN Company, and performed according to the instruction manual.

[0067] 2.2 Reverse transcription:

[0068] The reverse transcription kit (DRR047) from TAKARA company was used to reverse 1 μg RNA. Compared with the traditional reverse transcription kit, the kit added the step of removing genomic DNA, which can ensure the purity of RNA and the efficiency of amplification to the greatest extent. specificity. The reaction system and reaction c...

Embodiment 3

[0079] Example 3 Study on the effect of miRNA-3613 on the adhesion ability of lung squamous cell carcinoma cells

[0080] 1. miRNA synthesis

[0081] The control miRNA (miRNA-NC), miR-3613 was synthesized by Shanghai Gemma Company, wherein the sequence of miR-3613 is shown in SEQ ID NO.4.

[0082] 2. Cell transfection

[0083] 2.1 Cell culture: The method for culturing NCI-H596 cells is the same as in Example 2.

[0084] 3. Cell transfection

[0085] The NCI-H596 cells were divided into two groups, namely control miRNA-NC group and miRNA-3613 group. The two groups were transfected with miRNA-NC and miRNA-3613, respectively, using the transfection reagent LipofectamineTM2000 for transfection, and the transfection method was referred to the instructions. The working concentration of both miRNA-NC and miRNA-3613 was 5 μM. Cells in each group were collected 48 h after transfection for subsequent experiments.

[0086] 4. QPCR experiment

[0087] The steps of extraction of to...

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Abstract

The invention discloses an application of miR-3613 as a miRNA marker in distinguishing metastasis and non-metastasis of lung squamous carcinoma. Accordingly, the miR-3613 can be used for developing products for diagnosing lung squamous carcinoma metastasis and drugs for treating lung squamous carcinoma metastasis. The research result of the invention provides a theoretical basis for making individual-based treatment schemes by clinical physicians and can provide a new drug target for developing lung squamous carcinoma drugs.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to the use of miR-3613 in the diagnosis and treatment of lung squamous cell carcinoma metastasis. Background technique [0002] Non-small cell lung cancer (NSCLC) is a complex disease that seriously threatens human life and health. It can occur in almost all parts, tissues, and organs of the human body. The treatment of non-small cell lung cancer is a multidisciplinary comprehensive treatment, in which chemotherapy is one of the important means of treatment, but the drug resistance of non-small cell lung cancer and metastasis of non-small cell lung cancer are the reasons for the low overall survival rate of non-small cell lung cancer patients. reason. Lack of effective early diagnosis and treatment for non-small cell lung cancer metastasis and drug resistance is the bottleneck in the current prevention and treatment. [0003] miRNA is a 21-22nt RNA molecule edited by intracellular gene tra...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68A61K45/00A61P35/04
CPCA61K45/00C12Q1/6886C12Q2600/158C12Q2600/178
Inventor 杨承刚宋宏涛李曙光
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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