Use of microRNA-548k inhibitor in treatment and/or prevention of esophageal squamous cancer

1. The technology of microrna-548k and inhibitors, which can be applied in gene therapy, microbial measurement/testing, biochemical equipment and methods, etc., and can solve problems such as the research of unreported functions

Active Publication Date: 2015-09-02
CANCER INST & HOSPITAL CHINESE ACADEMY OF MEDICAL SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] microRNA-548k is located on chromosome 11 and contains 22 nucleotides. Its genbank accession number is: Gene ID: 100313770, and no research on its related functions has been reported in the literature

Method used

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  • Use of microRNA-548k inhibitor in treatment and/or prevention of esophageal squamous cancer
  • Use of microRNA-548k inhibitor in treatment and/or prevention of esophageal squamous cancer
  • Use of microRNA-548k inhibitor in treatment and/or prevention of esophageal squamous cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1: Method for detecting microRNA-548k gene expression

[0031] 1. Extraction of total RNA from sample-Trizol method to extract total RNA from tissues and cells

[0032] (1) Sample processing (bacteria and cells do not need to be ground);

[0033] Break the tissue sample about 1cm2 in aluminum foil, transfer it to the Eppendorf tube with steel balls, and grind it with a grinder (301 / s, 8min)

[0034] Note: This step should be operated in a low-temperature liquid nitrogen environment as much as possible, bacteria and cell samples do not need to be ground.

[0035] (2) Add 1ml Trizol to the ground Eppendorf tube, shake and mix;

[0036] (3) Move the mixed solution to a new Eppendorf tube, add 200μl chloroform, shake and mix;

[0037] (4) Centrifugation: 4°C, 12000rpm, 15min;

[0038] (5) Transfer the supernatant after centrifugation to a new Eppendorf tube, add 500 μl isopropanol, mix gently, and let it stand at room temperature for 15 minutes;

[0039] (6) Centrifugation: 4℃, 1...

Embodiment 2

[0068] Example 2: The expression level of microRNA-548k in human esophageal cancer cell lines

[0069] Real-time PCR was used to detect the expression of microRNA-548k in esophageal squamous cell carcinoma cell lines and normal esophageal cells (see figure 1 ). Among them, the abscissa represents normal cell lines (NE2 and H5E973) and human esophageal cancer cell lines. Compared with normal cell lines, the expression level of microRNA-548k in human esophageal cancer cell lines increased.

Embodiment 3

[0070] Example 3: The effect of changing the expression of microRNA-548k on cell growth

[0071] In this embodiment, the antisense nucleic acid complementary to the microRNA-548k sequence is used as an inhibitor; in the case of "mimicry", the expression level of microRNA-548k in the cell is increased by transfection with the same sequence as microRNA-548k. In the case of inhibitors and mimicry, the expression level of microRNA-548k in cells was down-regulated by about 2-11 times, or up-regulated by about 50,000 times. In the case of the miR-NC negative control, the microRNA sequence transfected into the nematode is used, because it is not homologous to humans and humans have no related mircroRNA, so it can be used as a negative control.

[0072] In KYSE30, KYSE150 and KYSE410 cell lines, the effect of up-regulation of microRNA-548k level on the growth curve of the cell line was tested. Count 2000 cells respectively and inoculate on 96E-Plate, and put the inoculated E-Plate into th...

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Abstract

The invention relates to the use of a microRNA-548k inhibitor in treatment and / or prevention of the esophageal squamous cancer. Further the invention relates to the microRNA-548k as a molecule marker used for diagnosing the esophageal squamous cancer.

Description

Technical field [0001] The present invention relates to the technical field of microRNA-548k inhibitors in the treatment and / or prevention of esophageal squamous cell carcinoma. In another aspect, the present invention relates to the technical field of microRNA-548k as a molecular marker for the diagnosis of esophageal squamous cell carcinoma. Background technique [0002] Esophageal cancer is a common gastrointestinal tumor. About 300,000 people die from esophageal cancer each year worldwide. It is the sixth leading cause of cancer deaths in the world, see Kamangar, F., GMDores, et al. (2006)." Patterns of cancer incidence, mortality, and prevalence across five continents: defining priorities to reduce cancer disparities in different geographic regions of the world. "J Clin Oncol24(14): 2137-2150. Approximately 70% of esophageal cancers worldwide occur in China, and 90% of esophageal cancers in China are esophageal squamous cell carcinomas. So far, the mechanism of occurrence ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61P35/00C12Q1/68
Inventor 詹启敏李林宋咏梅高志博欧云尉王俊
Owner CANCER INST & HOSPITAL CHINESE ACADEMY OF MEDICAL SCI
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