Oral squamous carcinoma cell strain as well as preparation method and applications thereof

A technology for oral squamous cell carcinoma and cancer cells, applied in the field of life medicine

Inactive Publication Date: 2017-02-15
SHANGHAI NINTH PEOPLES HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this ignores the influence of the immune system on tumor occurrence and metastasis; when using such cell lines and nude mouse animal models to screen or develop new drugs for the treatment of cancer, the influence of the immune system on the effect of anticancer drugs is ignored. Also ignores the impact of cancer drugs on immune system function

Method used

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  • Oral squamous carcinoma cell strain as well as preparation method and applications thereof
  • Oral squamous carcinoma cell strain as well as preparation method and applications thereof
  • Oral squamous carcinoma cell strain as well as preparation method and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] The mouse oral cancer oral cancer cell line is prepared according to the following method:

[0034] Step 1: Acquisition of mouse oral cancer tissue blocks: After drinking water to induce MAL gene knockout mice for 20 weeks with the conventional mouse oral cancer inducer 4NQO (80 μg / ml), the mucosa of the back of the tongue of a female mouse numbered 167# A cauliflower-shaped lesion was found, and the pathological diagnosis was oral squamous cell carcinoma, see figure 2 , use sterilized surgical instruments to quickly remove the tumor on the tip of the tongue, and use sterilized small scissors to quickly dissociate the tumor specimen, soak it in high-sugar DMEM solution containing 1% penicillin-streptomycin double antibody, and move it to a biological safety cabinet, containing double antibody The remaining tissue was obtained after repeatedly washing the tissue with sterile PBS buffer to remove impurities such as mucus and red blood cells;

[0035] Step 2: Tissue bloc...

Embodiment 2

[0040] Mice oral cancer cell line Mca-T-MAL of the present invention - / - detection and identification

[0041] 1. Morphological observation

[0042] Mainly observe the general morphology of cells, such as gross shape, nucleoplasmic ratio, chromatin and nucleolus size, number, etc., as well as the arrangement of cytoskeleton microfilaments and microtubules.

[0043] 1.1 Inverted optical microscope observation

[0044] The general morphology of cells, such as general shape, nuclear-to-cytoplasmic ratio, and adhesion characteristics, is mainly observed in the normal growth state.

[0045] Observing the mouse oral cancer cell line Mca-T-MAL of the present invention under an inverted optical phase contrast microscope - / - , it can be seen that the cells are polygonal or elliptical, grow in groups, have large nuclei, abundant cytoplasm, tight connections between cells, and grow attached to the plate. HE staining shows that the nucleus is round, oval or kidney-shaped, the staining...

Embodiment 3

[0084] Mice oral cancer cell line Mca-T-MAL of the present invention - / - Establishment of in vivo animal models

[0085] 6-week-old BALB / C nude mice were selected for the experiment, half male and half male, and all animal experiments were carried out in accordance with the operating guidelines for animal experiments approved by Shanghai Jiaotong University School of Medicine.

[0086] 1. Subcutaneous tumor formation in nude mice

[0087] Step 1: The experimental cells are divided into two groups, which are respectively the cell line Mca-T-MAL of the present invention - / - and unselected primary mouse oral carcinoma cells;

[0088] Step 2: After cell digestion and counting, wash 2 times with serum-free and antibiotic-free DMEM, and dilute the cells to 1×10 with serum-free and double-antibody-free DMEM. 7 cells / ml of cell suspension, kept on ice;

[0089] Step 3: select two injection points for each nude mouse, inject 100 μl of cell suspension subcutaneously on the outer sid...

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Abstract

The invention provides an oral squamous carcinoma cell strain as well as a preparation method and applications of the oral squamous carcinoma cell strain. The oral squamous carcinoma cell strain is derived from the MAL-/-C57BL/6 mouse oral carcinoma tissue induced by drinking water containing 4-nitroquinoline-1 oxide. Through primary culture on the tumor tissue, the tumour cell is subjected to continuous subculture in vitro and grows lively. Through the monoclonal screening method, the oral squamous carcinoma cell strain is obtained, the cell strain can be cultured continuously in vitro and grows lively, and the following biological characteristics of the cell are analyzed: cell proliferation, growth curve, cell cycle and apoptosis, plate colony and migration, and the in-vivo tumor formation and pulmonary metastasis capacity of the nude mouse. The result proves that the oral squamous carcinoma cell strain has the typical biological characteristics of the tumor cell, can be taken as the cell model for researching the generation, development and metastasis mechanism of oral carcinoma, and also can be taken as a tool for screening or developing anti-cancer drugs.

Description

technical field [0001] The invention belongs to the field of life medicine, and in particular relates to an oral squamous cell carcinoma cell line, its preparation method and related applications. Background technique [0002] Oral cancer is an important part of head and neck malignancies, and its incidence is increasing year by year. Among oral cancers, more than 90% are oral squamous cell carcinomas (OSCC). Oral squamous cell carcinoma refers to squamous cell carcinoma that occurs in the normal mucosal epithelium of the oral cavity, also known as oral mucosal cell carcinoma. Smoking, alcohol abuse, and a stimulating diet are considered major risk factors for oral squamous cell carcinoma. Since the oral cavity is located in the middle of the head and neck, and it is also the common starting point of the respiratory system and digestive system, malignant tumors in this area often directly affect the patient's important functions such as swallowing, chewing, breathing, spea...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09C12Q1/02A01K67/027C12R1/91
CPCC12N5/0693A01K67/0276A01K2217/075A01K2227/105A01K2267/0331C12N2503/02C12N2509/00G01N33/5011
Inventor 祁胜财陈万涛吴祥冰曹巍严明朱雪琴王晓宁
Owner SHANGHAI NINTH PEOPLES HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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