Molecular marker for esophageal cancer cell line migration phenotypes and application thereof

A technology of molecular markers and cancer cells, applied in the field of tumor molecular biology, can solve the problems of difficult migration of esophageal cancer cells, delayed disease progression, and procrastination

Active Publication Date: 2019-12-10
THE FIRST AFFILIATED HOSPITAL OF ZHENGZHOU UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, at present, there are few molecular markers related to judging the migration of early esophageal cancer cells, which makes it difficult to judge whether the migration of esophageal cancer cells in the early stage, thus delaying the disease.

Method used

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  • Molecular marker for esophageal cancer cell line migration phenotypes and application thereof
  • Molecular marker for esophageal cancer cell line migration phenotypes and application thereof
  • Molecular marker for esophageal cancer cell line migration phenotypes and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 Western blot detection of differential expression of RNF31 gene protein level

[0022] The cells involved in this example (EC9706) were purchased from the Cell Bank of Shanghai Chinese Academy of Sciences.

[0023] 1.1. Obtain transfected esophageal cancer cell lines:

[0024] siRNA—Small interfering RNA that causes the degradation of the mRNA of a target gene that matches its sequence. This results in a phenotype of gene silencing in cells or organisms.

[0025] In this example, NC, siRNF31#1 and siRNF31#2 were purchased from sigma company, and NC is a nonsense RNA sequence that does not cause any gene silencing phenomenon. The detailed sequence is as follows:

[0026] NC UUCUCCGAACGUGUCACGUTT ACGUGACACGUUCGGAGAATT

[0027] siRNF31#1 GCGAUUAUAUGGCUACACATT UGUGUAGCCAUAUAAUCGCTT

[0028] siRNF31#2 GGCGUGGUGUCAAGUUUAATT UUAAACUUGACACCACGCCTT

[0029] The siRNA or control was transfected into the esophageal cancer cell line EC9706 by liposome-mediated trans...

Embodiment 2

[0040] Example 2 Q-PCR detection of differential expression of RNF31 gene mRNA level

[0041] The cells involved in this example (EC9706) were purchased from the Cell Bank of Shanghai Chinese Academy of Sciences.

[0042] 2.1. Obtain transfected esophageal cancer cell lines:

[0043] The siRNA or control was transfected into the esophageal cancer cell line EC9706 by liposome-mediated transfection. The detailed steps of cell culture and transfection are as follows:

[0044] Culture and subculture of EC9706 cells: the cells were purchased from the Cell Bank of the Chinese Academy of Sciences, and the cells were placed in CO 2Cultivate in an incubator. After the cells are completely attached to the wall, discard the original culture, add fresh culture medium RPMI1640+10% FBS to continue the culture, observe the cell morphology and growth status under an inverted microscope every day, and wait until the cells are full of the culture bottle The bottom 90% or so began to be subcu...

Embodiment 3

[0058] Example 3 Differences in scratch experiments after RNF31 gene silencing

[0059] The cells involved in this example (EC9706) were purchased from the Cell Bank of Shanghai Chinese Academy of Sciences.

[0060] 3.1. Obtain transfected esophageal cancer cell lines:

[0061] The siRNA or control was transfected into the esophageal cancer cell line EC9706 by liposome-mediated transfection. The detailed steps of cell culture and transfection are as follows:

[0062] Culture and subculture of EC9706 cells: the cells were purchased from the Cell Bank of the Chinese Academy of Sciences, and the cells were placed in CO 2 Cultivate in an incubator. After the cells are completely attached to the wall, discard the original culture, add fresh culture medium RPMI1640+10% FBS to continue the culture, observe the cell morphology and growth status under an inverted microscope every day, and wait until the cells are full of the culture bottle The bottom 90% or so began to be subculture...

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Abstract

The invention belongs to the technical field of tumor molecular biology and particularly relates to a molecular marker for esophageal cancer cell line migration phenotypes and application thereof. RNF31 is an E3 ubiquitin ligase. The invention discovers that expression of an RNF31 gene is related to the migration capability of an esophageal cancer cell line for the first time, and the intensity ofthe migration capability of the tested esophagus cancer cell line can be judged by detecting expression of the RNF31 in the tested esophageal cancer cell line; tested esophagus cancer cells are transfected with a target gene siRNA and does not contain the target gene siRNA respectively, and the siRNA silence efficiency is detected by utilizing western blot and a q-PCR technology; for judgment onmigration of the esophagus cancer cell line, the migration capacity of the cells can be accurately reflected through scratches and transwell experiments; and the migration phenotype of an esophagus cancer can be rapidly judged according to detection of RNF31 proteins and the mRNA content of the tested esophagus cancer cell line.

Description

technical field [0001] The invention belongs to the technical field of tumor molecular biology, and in particular relates to a molecular marker of migration phenotype of esophageal cancer cell line and application thereof. Background technique [0002] China is one of the countries with the highest morbidity and mortality rates of esophageal cancer in the world. The Taihang Mountains, including parts of Henan, Hebei, and Shanxi provinces, is a traditional high-incidence area for esophageal cancer in my country. However, corresponding to the grim epidemiological situation of esophageal cancer is the lack of early diagnosis of esophageal cancer. Most patients are already in the advanced stage of esophageal cancer when they see a doctor. Although there are many improvements in treatment methods, the 5-year survival of patients The overall rate is still hovering around 15%, especially for advanced esophageal cancer, the effect is not ideal. Early detection and treatment of eso...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/158
Inventor 王立东李秀敏侯婧晗庞丹卢奎张爱佳王素杰高灿吕本洁闫子怡周孝峰王伟隆陈贞
Owner THE FIRST AFFILIATED HOSPITAL OF ZHENGZHOU UNIV
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