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Chinese lung squamous carcinoma cell line and application thereof

A technology of Chinese people and cancer cells, applied in the field of tumor biology, can solve problems such as ineffectiveness

Active Publication Date: 2020-10-02
HUADONG HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most lung squamous cell carcinomas do not have k-ras and other gene mutations, and targeted drugs have little effect on them. Traditional chemotherapy and radiotherapy are still the main treatment for this type of lung cancer

Method used

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  • Chinese lung squamous carcinoma cell line and application thereof
  • Chinese lung squamous carcinoma cell line and application thereof
  • Chinese lung squamous carcinoma cell line and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Isolation and processing of primary tumor cells

[0027] Primary tumor cells were isolated and cultured in a sterile environment from fresh tumor tissue blocks obtained from the animal room.

[0028] 1. First put the tumor tissue block into a 50ml centrifuge tube, shake and wash 5 times with PBS containing 10 times the double antibody until there is no blood. Place the tissue block in a Petri dish and remove excess tissue, such as fat or dead tissue.

[0029] 2. Move the tissue to another plate, and use a cross scalpel to cut it into about 1mm 3 The size of the organization block. Take a 70um filter and place it on a 50ml centrifuge tube. Gently pressurize the core of the syringe to force the tissue through the mesh into the culture medium, draw the culture medium and blow it through the mesh to flush the cells down.

[0030] 3. Add 5ml of trypsin / collagenase II to the culture dish of the remaining tissue pieces, digest in a 37°C incubator for 2 hours, and pipette e...

Embodiment 2

[0044] Example 2. Detection and identification of Chinese lung squamous cell carcinoma cell line SAHP1

[0045] 1. Cytomorphological observation of primary tumor cell lines

[0046] Under the light microscope, most of the tumor cells are spindle-shaped and oval-shaped, and a few are polygonal. Cells in the logarithmic growth phase spread well. SAHP1 spread randomly, and most of them were oval epithelioid cells, polygonal, and more regular in shape. Such as figure 1 shown.

[0047] 2. Immunofluorescence detection of cell marker proteins

[0048] 1) After trypsinizing the cells, suspend them in 1mL of complete culture medium, pipette fully to form a single-cell suspension, and perform cell counting. Insert 5×10 into 24-well plate as needed 4 cells. Before adding the cell suspension, add a small amount of medium to each well to make the slide and the culture dish stick together to reduce the entry of cells and prevent the slide from floating.

[0049] 2) Wash the slide with ...

Embodiment 3

[0056] Example 3. Karyotype analysis

[0057] Karyotype analysis to detect the characteristics of the karyotype and the number of chromosomes

[0058] 1) Add colchicine to the cells in the logarithmic growth phase, so that the final concentration of colchicine is 0.1ug / ml, and continue to culture for 3h;

[0059] 2) Digest the cells and transfer them to a 10ml pointed centrifuge tube, centrifuge at 1000rpm for 10min, remove the supernatant, add 0.075mol / L KC110ml, incubate at 37°C for 30min, then add fresh fixative solution (glacial acetic acid:methanol) to the centrifuge tube =3:1) 1ml, mix well, centrifuge at 1000rpm for 10min, discard the supernatant;

[0060] 3) Slowly add 10ml of fresh fixative solution, gently blow evenly, fix at room temperature for 20min, centrifuge, remove the supernatant, and fix again; centrifuge at 1000rpm for 10min, discard the supernatant;

[0061] 4) Add 250ul of fresh fixative solution, blow gently, drop a drop of suspension solution at a hei...

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Abstract

The invention provides a Chinese lung squamous carcinoma cell line which is named as Chinese lung squamous carcinoma cell line SAHP1, and the preservation number of the Chinese lung squamous carcinomacell line SAHP1 is CCTCC NO: C2019266. The cell line provided by the invention has similar protein expression with a primary tumor tissue, also has chromosomal abnormal karyotype, and has the tumor characteristic of an in-vitro tumor formation capability. The cell line provided by the invention can be subjected to in-vitro continuous subculture to keep the characteristics of lung squamous carcinoma tumor cells unchanged, and is suitable for being used as a cell material for researching and developing lung squamous carcinoma related immune treatment, diagnosis and drugs. The cell line disclosed by the invention contains Kras-G12V mutation, and can be used as a tool cell for researching a targeted drug taking the Kras-G12V mutation as a target.

Description

technical field [0001] The invention relates to the technical field of tumor biology, in particular to a Chinese lung squamous cell carcinoma cell line and its application. Background technique [0002] Lung cancer is one of the tumors with the highest mortality rate in the world, accounting for 19.4% of the deaths caused by tumors. In Europe, with the reduction of the number of smoking in China and the increase of female smokers, the incidence rate of men is decreasing year by year, and the incidence rate of women is increasing year by year. Among them, 80%-85% are non-small cell lung cancer, and the 5-year survival period is low at 11%. The epidemiological study of lung cancer in Asia in 2014 showed that 43.4% of lung cancers were female, the average age of onset was 60 years old, and 52.6% had no smoking history. Lung cancer is mainly divided into non-small cell lung cancer and small cell lung cancer. Non-small cell lung cancer is mainly divided into adenocarcinoma, sq...

Claims

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Application Information

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IPC IPC(8): C12N5/09C12Q1/02A01K67/027C12R1/91
CPCC12N5/0693C12N5/0688G01N33/5011G01N33/5044A01K67/0271C12N2503/02G01N2500/10A01K2207/15A01K2227/105A01K2267/0331
Inventor 高文路丽明武韫张娜潘常青唐东方汪凯
Owner HUADONG HOSPITAL
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