Use of micro-RNA molecule miR491-5p in treatment and/or diagnosis and/or prognosis of pancreatic cancer

A technology for pancreatic cancer and its use, which is applied in the fields of biomedicine, gene therapy and/or gene diagnosis, and can solve problems such as the influence of pancreatic cancer treatment and diagnosis and prognosis, and the undetected hsa-miR491-5p pancreatic cancer correlation.

Active Publication Date: 2014-06-18
THE INST OF BASIC MEDICAL SCI OF CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, the association of hsa-miR491-5p (miR491-5p for short) with pancreatic carcinogenesis was not detected during these large-scale miRNA microarray screens
The inventor's research shows that miR49-5p is highly expressed in normal human pancr

Method used

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  • Use of micro-RNA molecule miR491-5p in treatment and/or diagnosis and/or prognosis of pancreatic cancer
  • Use of micro-RNA molecule miR491-5p in treatment and/or diagnosis and/or prognosis of pancreatic cancer
  • Use of micro-RNA molecule miR491-5p in treatment and/or diagnosis and/or prognosis of pancreatic cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Example 1. Tissue cell expression profile analysis and detection of miR491-5p

[0060] Do the following:

[0061] Amplify human embryonic kidney cell line 293, human liver cancer cell line HepG2, human lung cancer cell line A549, human pancreatic cancer cell line Cap-1 and SW1990 in a cell culture dish; collect at least 5×10 cells for each 6 Treat with 1 mL TRIzol, add 150 μl chloroform, mix and centrifuge to get the supernatant; add an equal volume of isopropanol to the supernatant to precipitate, and obtain the total cellular RNA;

[0062] Total RNA was extracted from normal adult tissues (human lung, human kidney, human brain, human liver, human spleen and human pancreas) and normal human embryonic tissues (human embryonic liver, human embryonic spleen and human embryonic pancreas);

[0063] 100ng of total RNA was used as a template for poly(A) RT-qPCR analysis (Zhang, J. et al., 2008). The instrument used was the iQ5 real-time PCR detection system (Bio-Rad Laborato...

Embodiment 2

[0073] Example 2. Detection of correlation between miR491-5p and occurrence of pancreatic cancer

[0074] Extract the total RNA of pancreatic cancer cell line Cap-1, pancreatic cancer cell line SW1990, pancreatic cancer cell line Mia, and pancreatic cancer cell line AsPC-1;

[0075] Obtain the total RNA of normal human islet β cells and the total RNA of normal human stellate cells;

[0076] Detection was performed with poly(A) RT-qPCR.

[0077] result:

[0078] miR491-5p is highly expressed in normal human pancreatic tissue, and its expression level is also very low in pancreatic cancer cell lines Mia and AsPC-1 cells. The expression level of miR491-5p in pancreatic islet β cells and pancreatic stellate cells is also maintained at a low level. Level( figure 2 ). It shows that the abnormal expression of miR491-5p is probably related to the occurrence of pancreatic cancer.

Embodiment 3

[0079] Example 3. Construction and detection of miR-491-5p expression vector

[0080] From the miRNA database (http: / / microrna.sanger.ac.uk / sequences / ), find the position and specific sequence information of miR-491-5p on the genome (SEQ ID No.2);

[0081] According to the genome sequence, determine the specific position of pri-miR491-5p;

[0082] Design specific primers in the 500-800bp region upstream and downstream of the pri-miR-491-5p site: SEQ ID No.8: 5′-ttagatctacagaagctgcacacataca-3′ (forward primer)

[0083] SEQ ID No.9: 5'-ttgtcgactatctcaactgctgccatca-3' (reverse primer);

[0084] PCR amplification of the target sequence was performed using the genomic DNA of HEK293T cells as a template. The amplification conditions were denaturation at 94°C for 5 minutes, followed by 94°C for 30 seconds, 50°C for 30 seconds, and 72°C for 90 seconds, a total of 35 cycles, and finally 72°C 10 minutes;

[0085] Perform 1% agarose gel electrophoresis detection on PCR amplification p...

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Abstract

The present invention relates to a use of micro-RNA molecule miR491-5p in treatment and/or diagnosis and/or prognosis of pancreatic cancer, and particularly discloses a use of the micro-RNA molecule miR491-5p in preparation of pancreatic cancer treatment drugs. The present invention further discloses a use of the micro-RNA molecule miR491-5p in preparation of an apparatus for diagnosis and/or prognosis of pancreatic cancer. According to the present invention, the miR491-5p presents differential expression between the normal pancreas and the pancreatic cancer tissue, can be used as the pancreatic cancer diagnosis marker, and can further be used as the active ingredient so as to be used for preparing the pancreatic cancer treatment drug.

Description

technical field [0001] The present invention relates to the field of biomedicine, and relates to the field of gene therapy and / or gene diagnosis; in particular, it relates to the use of microRNA molecules in the preparation of drugs for treating pancreatic cancer and the use of microRNA molecules in the preparation of drugs for diagnosis and / or prognosis of pancreatic cancer. Use in a device for cancer; more specifically, use of miR491-5p in the preparation of a drug for treating pancreatic cancer and use of miR491-5p in the preparation of a device for diagnosis and / or prognosis of pancreatic cancer. Background technique [0002] MicroRNA (microRNA or miRNA) is a new type of gene regulator that can actively participate in physiological and pathological processes. This type of RNA is a class of small non-coding RNAs approximately 21-25 nucleotides (nt) in length, whose function is to regulate the expression of target genes at the post-transcriptional level (Bartel, D.P., 2004...

Claims

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Application Information

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IPC IPC(8): A61K48/00C12Q1/68A61P35/00
Inventor 刘力郭蓉
Owner THE INST OF BASIC MEDICAL SCI OF CHINESE ACAD OF MEDICAL SCI
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