miR-506 targeting PENK gene of triple negative breast cancer cells and application of miR-506

A triple-negative breast cancer, mir-506 technology, applied in the field of molecular biology and oncology, can solve the problem of improving triple-negative breast cancer cells

Pending Publication Date: 2020-02-07
NORTHWESTERN POLYTECHNICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no research on increasing the expression of penk in triple-negative breast cancer cells by targeting the miRNA pathway. Therefore, it is important to explore the pathogenesis of penk-related TNBC and find the corresponding miRNA as the molecular target for the treatment of TNBC. significance

Method used

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  • miR-506 targeting PENK gene of triple negative breast cancer cells and application of miR-506
  • miR-506 targeting PENK gene of triple negative breast cancer cells and application of miR-506
  • miR-506 targeting PENK gene of triple negative breast cancer cells and application of miR-506

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1 Discovery and verification of miR-506 targeting penk in triple-negative breast cancer.

[0059] Through research, the inventor found that the expression of Penk was significantly down-regulated in multiple tissue samples of triple-negative breast cancer patients. In order to further explore the mechanism affecting the expression of Penk, the inventor searched for proteins or RNAs that may interact with Penk through websites or software. miRNA is a single-stranded RNA molecule of about 22nt, which does not code for protein. miRNA mainly binds to the 3' untranslated region (3'-UTR) of the target mRNA to inhibit mRNA translation or degrade mRNA, thereby reducing the expression of the corresponding protein, thereby affecting cell proliferation, metastasis, apoptosis, etc. a series of life activities.

[0060] 1. Prediction and verification of Penk-related miRNAs

[0061] 1) Using the database (miRWalk 2.0 (http: / / zmf.umm.uni-heidelberg.de / apps / zmf / mirwalk2 / index...

Embodiment 2

[0102] Example 2 Detection of the influence of miR-506 on the biological functions of triple-negative breast cancer cells by targeting penk

[0103] 1) The effect of miR-506 on the proliferation of MDA-MB-231 cells

[0104] a. Use a 96-well plate to plate, and the amount of cells inoculated in each well is 1×10 3 indivual. Three groups were set up, namely NC group, miR-506 mimic group, and miR-506 inhibitor group, with six replicates in each group. The next day, the cells in each well were mixed with 0.2 μL 2000, 0.5 μL RNA was used for transfection.

[0105] b. On the 3rd, 4th, and 5th day after transfection, the cell viability was detected by MTT method. Each well was replaced with 100 μL of fresh medium, and 10 μL of MTT solution was added in the dark, and incubated at 37°C for 4 h. Aspirate the mixed solution in the wells, add 100 μL DMS0 solution to each well, shake for 10 min, obtain the absorbance values ​​of the mixed solution at the wavelengths of 570 nm and 650...

Embodiment 3

[0133] Example 3 Effect of miR-506 Local Sustained Release System Drugs on Nude Mouse Breast Cancer Model

[0134] 1) Construction of miRNA local sustained release system

[0135] Take 16 mg of gelatin nano-microspheres and irradiate them with ultraviolet light for 30 minutes to sterilize them. Take 32.5 μL 2000 was added to 45 μL Opti-MEM, and the concentration of RNA was adjusted to 1 μg / μL, and 10 μL RNA was added to 32.5 μL Opti-MEM, flicked and mixed, and stood for 5 minutes. Add RNA mix to 2000, stand at room temperature for 20 minutes. Mix the mixture with the microspheres, and let stand at 4°C for 1 hour to mix well and set aside.

[0136] 2) Establishment of breast cancer model in nude mice

[0137] a. Cultivate a sufficient amount of MDA-MB-231 cells and collect them in a 50mL centrifuge tube. Dilute Matrigel to 3 mg / mL with serum-free L-15 medium, take 3 mL of Matrigel mixture and resuspend the cells so that the density of each 100 μL cell suspension is 2×10...

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Abstract

The invention relates to a miR-506 targeting PENK gene of triple negative breast cancer cells and application of the miR-506. By screening miRWalk2.0, miRBase, TargetScanHuman 7.0 databases and detecting tissue samples of triple negative breast cancer patients by using a qRT-PCR technology, it is confirmed that the expression of miR-506 in the triple negative breast cancer is decreased significantly, and is averagely down-regulated by 88.95%, a dual-luciferase reporter experiment confirms that miR-506 can directly interact with Penk, and it is indicated that miR-506 can target PENK. Accordingto the miR-506 targeting the PENK gene of the triple negative breast cancer cells and the application of the miR-506, it is proved that miR-506 affects the proliferation, metastasis and invasion of triple negative breast cancer cells by increasing the content of penk in a targeting mode; the effectiveness of miR-506 targeting penk in inhibiting the progression of the triple negative breast canceris detected through a nude mouse breast cancer model; and the results show that the miR-506 can inhibit the development of the triple negative breast cancer through targeting penk.

Description

technical field [0001] The invention belongs to the field of molecular biology and oncology, and specifically relates to miR-506 targeting triple-negative breast cancer cell PENK gene and application thereof. Background technique [0002] Breast cancer is the most common cancer among women worldwide. Although the mammary gland is not an essential tissue for human life activities, the cells of the mammary gland tissue are loosely connected after canceration and are easy to fall off, and the shed cancer cells will transfer to the whole body along with the blood circulation or lymphatic circulation system, causing canceration in other tissues and ultimately threatening life . Triple negative breast cancer (TNBC) is a subtype of breast cancer whose expression of progesterone receptor, estrogen receptor, and human epidermal growth factor receptor 2 is negative. Triple-negative breast cancer accounts for 10.0% to 20.8% of all breast cancers. It has special clinicopathological fe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113A61P35/00A01K67/027
CPCC12N15/113A61P35/00A01K67/0275C12N2310/141A01K2207/05A01K2217/058A01K2227/105A01K2267/0331
Inventor 张辰艳刘杰刘文婧尹大川杨长青刘新利董凯赵士淇
Owner NORTHWESTERN POLYTECHNICAL UNIV
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