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Male esophageal carcinoma cell system and application thereof

A technology for cancer cells and esophagus, applied to male esophageal cancer cell lines and its application fields, can solve the problems of unreported cytokeratin cytokeratin, easy to be polluted, and inability to carry out in-depth research on esophageal cancer

Inactive Publication Date: 2018-11-27
THE FIRST HOSPITAL OF HEBEI MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are few esophageal squamous cell lines established in China, and they are easily contaminated, so that the research on esophageal cancer cannot be carried out in depth. Moreover, none of the above esophageal cancer cell lines have been reported to express cytokeratin (CK) and cytokeratin (CK5). / 6) Features

Method used

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  • Male esophageal carcinoma cell system and application thereof
  • Male esophageal carcinoma cell system and application thereof
  • Male esophageal carcinoma cell system and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Cell Culture

[0028] (1) Primary culture: from a patient with esophageal cancer (Mongolian, male, 57 years old, the tumor was located in the middle thoracic esophagus, postoperative pathology showed grade I medullary squamous cell carcinoma of the esophagus, invading the whole layer, clinically No cancer, no lymph node metastasis, pT3N0M0, the patient was a patient in the First Hospital of Hebei Medical University who signed the informed consent) obtained 3-5 fresh clinical esophageal cancer endoscopic biopsy specimens, each about 0.2×0.2 ×0.2cm 3 Size, wash the tumor tissue pieces with PBS buffer (containing 1000U / ml penicillin G, 1000μg / ml streptomycin) 3 times, remove connective tissue and necrotic tissue, soak for 5-10min; soak the tissue pieces in 10% fetal bovine In the culture dish of serum 1640 medium, use a sterile scalpel to chop the tissue pieces in the culture dish, suck up the culture medium in the culture dish, put the chopped tissue pieces int...

Embodiment 2

[0032] Embodiment 2 Morphological observation

[0033] The male esophageal cancer cell line cultivated in Example 1 was placed under an inverted microscope, and photographed in bright field, it can be seen that the cells grow vigorously, the background is clear, and the cells are irregular polygons (see figure 1 ), conforming to the characteristics of epithelioid cells, named human esophageal cancer cell EC-1702, and was deposited in the China Center for Type Culture Collection on October 11, 2017, with the preservation number CCTCC NO: C2017202, the human esophageal cancer cell EC-1702 The -1702 doubling time is about 130 hours.

[0034] Smear the digested human esophageal cancer cell EC-1702 suspension, fix with 95% alcohol for 1 hour, wash with water, hematoxylin for 15 seconds, wash with water, acidify with hydrochloric acid alcohol, wash with water, turn blue with ammonia water, wash with eosin for 2 minutes, Washed with water, dehydrated with graded alcohol, mounted wi...

Embodiment 3

[0035] Example 3 tumor cell immunohistochemistry:

[0036]Human esophageal cancer cells EC-1702 were digested and centrifuged, and the centrifuged cell suspension was made into a cell smear, fixed with 95% alcohol for 1 hour, soaked in PBS buffer for later use. Boil the citric acid repair solution for 20 minutes and place it at room temperature, rinse with PBS for 5 minutes × 2 times; add about 50 μl of primary antibody (CK and CK5 / 6) dropwise, and freeze overnight at 4 ° C, and place it at room temperature the next day with PBS buffer for 5 minutes × 2 times , add biotinylated secondary antibody working solution dropwise, incubate at room temperature for 30min, wash with PBS, 5min×2 times; add horseradish enzyme-labeled streptavidin working solution dropwise, incubate at room temperature for 30min, wash with PBS, 5min×2 times; prepare fresh DAB solution, develop color for 5 minutes, observe under the microscope, stop color development with water, counterstain with hematoxylin...

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Abstract

The invention discloses a male esophageal carcinoma cell system and an application thereof. The male esophageal carcinoma cell system is preserved in China Center for Type Culture Collection (CCTCC),the preservation number of which is CCTCC NO: C2017202. The male esophageal carcinoma cell system can be used for finishing various molecular biological experiments. As the primary cell experimental result is more reliable than a cell strain experimental result and is closer to a clinical individual experimental result, the male esophageal carcinoma cell system is established to have important meaning in studying morbidity and treatment of Mongoloid (yellow race) esophageal carcinoma.

Description

technical field [0001] The present invention relates to the field of human esophageal cancer cell lines, in particular to male esophageal cancer cell lines and applications thereof. Background technique [0002] Esophageal cancer is a common malignant tumor of the digestive system. China is a country with the highest incidence and mortality rate of esophageal cancer in the world. According to GLOBOCAN data in 2012, there were 456,000 new cases of esophageal cancer in the world, and 223,000 cases occurred in China, accounting for 48.9%. There were 400,000 deaths from esophageal cancer in the world, and 197,000 cases died in China, accounting for 49.3%. The prevention and control situation is severe. There are two main subtypes of esophageal cancer in my country, squamous cell carcinoma and adenocarcinoma, of which squamous cell carcinoma accounts for about 90%. [0003] At present, the pathogenesis of esophageal squamous cell carcinoma has not been fully elucidated. In order...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09C12R1/91
CPCC12N5/0693
Inventor 于卫芳韩双双王嘉朱振龙刘德华赵红艳
Owner THE FIRST HOSPITAL OF HEBEI MEDICAL UNIV