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A screening method for marker proteins of lgien and hgien before esophageal squamous cell carcinoma

A screening method and protein technology, applied in the field of molecular biology, can solve the problems of difficult to clean tissue, no experimental method for protein extraction and identification, and inability to find precancerous molecular markers, and achieve the effect of simple experimental process.

Inactive Publication Date: 2019-01-22
FOURTH MILITARY MEDICAL UNIVERSITY
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Problems solved by technology

However, little research has been done on the differential proteins in different stages of precancerous lesions
This situation is mainly caused by the following reasons: 1. There are still certain technical difficulties in obtaining pure paraffin tissue samples
2. There is no general experimental method for the extraction and identification of proteins in pure paraffin-embedded tissues
3. It is not easy to obtain pure tissue, which directly leads to a rough distinction between cancer and paracancerous tissue, and it is impossible to really study the differential proteins of precancerous stages, and it is impossible to find general precancerous molecular markers. Effective primary prevention of ESCC

Method used

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  • A screening method for marker proteins of lgien and hgien before esophageal squamous cell carcinoma
  • A screening method for marker proteins of lgien and hgien before esophageal squamous cell carcinoma
  • A screening method for marker proteins of lgien and hgien before esophageal squamous cell carcinoma

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Embodiment 1

[0062] Preparation of membrane slides (PEN slides) for laser microdissection

[0063] 1. Select four post-ESD FFPE tissues of patients with squamous cell carcinoma of the esophagus, numbered as patient A's FFPE tissue, patient B's FFPE tissue, patient C's FFPE tissue, and patient D's FFPE tissue. , the FFPE tissue of patient B, the FFPE tissue of patient C and the FFPE tissue of patient D were prepared for HE slide specimens and PEN slide specimens respectively,

[0064] The preparation of HE slide specimens includes: cutting the FFPE tissue into 3-4μm thick tissue slices, hanging on a common slide and performing HE staining.

[0065] The preparation of PEN slide specimens includes: cutting the same FFPE tissue continuously at the same time, each tissue is 10 μm thick, spreading the slices in distilled water at 45°C, and picking up the slices in PEN membrane slides.

[0066] Ensure that the tissue samples on HE slides and PEN slides made from FFPE tissue from the same patie...

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Abstract

The invention discloses a method for screening labeled protein of esophageal squamous cell carcinoma front LGIEN and HGIEN. The method includes the steps that a formalin fixation paraffin embedding sample (FFPE sample) obtained after an esophageal squamous cell carcinoma ESD operation is manufactured into an HE slide and a PEN slide, the FFPE sample on the HE slide is subjected to pathological grading labeling, the FFPE sample on the PEN slide is subjected to dewaxing hydration, and grading areas on the HE slide obtained after grading labeling are rubbed on the PEN slide obtained after dewaxing hydration; the rubbed PEN slide is subjected to cutting separation under a laser microscopic cutter according to rubbing areas, tissue obtained after cutting separation is sequentially subjected to protein extraction and determination and protein purification and enzymolysis, and protein of all the grading regions is obtained; the protein of all the grading regions is subjected to mass spectrometry through a four-level rod rail ion trap mass spectrum, and the mass spectrometry result is subjected to label-free library search analysis through MaxQuant software environment (version 1.4.3.9) software, and the comparison result of the protein and the protein screening result are obtained.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to a method for screening marker proteins of pre-esophageal squamous cell carcinoma LGIEN and HGIEN. Background technique [0002] Esophageal squamous cell carcinoma (ESCC) is a malignant tumor with high incidence in my country. The development of ESCC is the result of a multi-step process. It develops from normal squamous epithelium to intraepithelial neoplasia and finally to invasive carcinoma. . In 2000, the World Health Organization divided intraepithelial neoplasia into two categories: low-grade intraepithelial neoplasia (LGIEN) and high-grade intraepithelial neoplasia (HGIEN). Studies have shown that compared with normal esophageal squamous epithelium, the risks of LGIEN and HGIEN developing into squamous cell carcinoma are 10 times and 28 times higher respectively. Identification will provide opportunities for primary prevention and pharmacological treatment of E...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02G01N30/06G01N33/68G01N33/574G01N33/531
CPCG01N30/02G01N30/06G01N33/531G01N33/57484G01N33/68G01N33/6848G01N33/6893G01N2030/027G01N2030/062
Inventor 储玮李増山贾娜贾旭春陈玲胡思隽张莹肖黎明郭勇
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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