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Application of miR-193a-3p

A technology of 1.mir-193a-3p, 2.mir-193a-3p, applied in the application field of miR-193a-3p, can solve problems such as lack of explanation

Active Publication Date: 2017-12-01
NANJING BIOHENG BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Whether the treatment and early prevention of diabetes can slow down or prevent the occurrence of pancreatic cancer, the existing research work has not explained clearly

Method used

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  • Application of miR-193a-3p
  • Application of miR-193a-3p
  • Application of miR-193a-3p

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Effect of miR-193a-3p on migration and metastasis of pancreatic cancer cells

[0061] Transfected or infected with pSIF-miR-193a-3p (miR-193a-3p) or pSIF-H1 (CtrlRNA) in BxPC-3 cells, scratch healing experiments, Trans-well cell migration experiments, Trans- well Matrigel invasion experiments, 3D cell culture experiments and bioimaging experiments:

[0062] Scratch Healing Experiment

[0063] Firstly, coordinate marking lines were drawn on the reverse side of the cell 6-well culture plate, and then the cells of different experiments and control groups were planted in the 6-well culture plate, and cultured with DMEM or DMEM / F12 complete medium. After the cell density reached 90%, the cells were used only The medium containing 0.2% serum concentration was cultured overnight. When the cell density reached 100, a 10ul pipette tip was used for scratching operation, and then photographed with an inverted microscope at different time points to observe the changes in migration...

Embodiment 2

[0090] miR-193a-3p can target DVL3 gene expression

[0091] Since the expression of miR-193a-3p in normal pancreatic tissue is significantly higher than that in pancreatic cancer tissue, miR-193a-3p may play an important tumor suppressor role in the development of pancreatic cancer. The inventors used the bioinformatics analysis software miRwalk2.0 software to predict the potential target genes of miR-193a-3p. We found Dishevelled3 (DVL3) as a potential target gene of miR-193a-3p ( Figure 7 -A, on). To verify whether miR-193a-3p directly targets and regulates DVL3, we first constructed a luciferase reporter gene plasmid containing wild-type DVL3 3′UTR ( Figure 7 -A, bottom), and the reporter gene plasmid DVL3 3′UTR-Mut plasmid containing miR-193a-3p seed sequence (7nt) deletion mutation ( Figure 7 -A, next). A dual-luciferase reporter assay was performed.

[0092] Dual Fluorescent Reporter Gene Assays

[0093] The predicted miRNA target gene 3'UTR was cloned into the ...

Embodiment 3

[0096] Effect of miR-193a-3p on migration and metastasis of insulin-treated pancreatic cancer cells

[0097] Pancreatic cancer cell line MIA PaCa-2 was treated with insulin (1 μM), insulin+miR-193a-3p, insulin+control RNA (Ctrl RNA). Insulin treatment of pancreatic cancer cell line MIA PaCa-2 with a low degree of malignancy, MTT cell proliferation and Trans-well cell migration analysis showed that the proliferation, migration and invasion abilities of MIA PaCa-2 cells were improved to varying degrees ( Figure 8 A-D). Since pancreatic cancer cells have strong tumor stem cell characteristics, we further analyzed the effect of insulin on the stemness of pancreatic cancer cells. Analysis of pancreatic cancer stem cell marker CD133 by flow cytometry found that the number of CD133-positive pancreatic cancer cells increased after insulin treatment ( Figure 8 -F); These experimental results show that insulin can significantly improve the migration and tumor formation abilities of ...

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Abstract

The invention belongs to the field of genetic engineering, and particularly relates to application of miR-193a-3p. The miR-193a-3p serves as a cancer suppressor gene in a pancreatic cancer cell to control migration and growing of the cancel cell, the miR-193a-3p can antagonist the promoting ability of insulin to proliferation, migration and invasion of the pancreatic cancer cell and a tumor stem cell, promotion and adjustment of the insulin to proliferation and migration of the pancreatic cancer cell by inhibiting the miR-193a-3p, and the application of the miR-193a-3p for preparing drugs for treating a pancreatic cancer and drugs for inhibiting the influence of the insulin on the pancreatic cancer is provided; and meanwhile the miR-193a-3p further can be used for diagnosing a marker indicating whether the insulin can be used for treating diabetic patients accompanied with the pancreatic cancer and preparing devices for judging whether the diabetic patients accompanied with the pancreatic cancer are suitable for being treated by adopting the insulin, and the basis can be provided by a judging result for a therapeutic schedule of the diabetic patients accompanied with the pancreatic cancer.

Description

technical field [0001] The invention belongs to the field of genetic engineering, in particular to the application of miR-193a-3p. Background technique [0002] Non-coding RNA refers to RNA that is transcribed by the genome but not translated into protein. Among them, microRNA (miRNA) is a type of small molecule non-coding RNA with evolutionary conservation, which has been confirmed to play an important role in cancer, cardiovascular, diabetes and other diseases ; miRNA has been shown to regulate the occurrence of cancer in different cancers and can be used as a potential clinical therapeutic target [0003] Pancreatic cancer is the cancer with the highest mortality rate. The 2-year survival rate is less than 20%, and the 5-year survival rate is only 2%. It is often called the "invisible killer", mainly because pancreatic cancer does not show any symptoms in the early stage, and often diagnosed pancreatic cancer patients are already in the advanced stage of cancer, so conve...

Claims

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Application Information

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IPC IPC(8): A61K31/7105A61K48/00A61P35/00C12Q1/68
CPCA61K31/7105C12Q1/6883C12Q2600/156C12Q2600/178
Inventor 贺小宏王延宾袁鹏王鑫张承武李林
Owner NANJING BIOHENG BIOTECH CO LTD
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