CN106011068A - Esophageal cancer cell line and application thereof - Google Patents
Esophageal cancer cell line and application thereof Download PDFInfo
- Publication number
- CN106011068A CN106011068A CN201610585742.4A CN201610585742A CN106011068A CN 106011068 A CN106011068 A CN 106011068A CN 201610585742 A CN201610585742 A CN 201610585742A CN 106011068 A CN106011068 A CN 106011068A
- Authority
- CN
- China
- Prior art keywords
- cell
- esophageal
- cell line
- cancer cell
- application
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 206010030155 Oesophageal carcinoma Diseases 0.000 title claims abstract description 60
- 208000000461 Esophageal Neoplasms Diseases 0.000 title claims abstract description 23
- 201000004101 esophageal cancer Diseases 0.000 title claims abstract description 23
- 206010028980 Neoplasm Diseases 0.000 claims abstract description 28
- 238000001514 detection method Methods 0.000 claims abstract description 14
- 208000017897 Carcinoma of esophagus Diseases 0.000 claims description 37
- 201000005619 esophageal carcinoma Diseases 0.000 claims description 37
- 208000007276 esophageal squamous cell carcinoma Diseases 0.000 claims description 32
- 238000011282 treatment Methods 0.000 claims description 8
- 238000011580 nude mouse model Methods 0.000 claims description 7
- 238000011160 research Methods 0.000 claims description 6
- 239000003814 drug Substances 0.000 claims description 5
- 241000124008 Mammalia Species 0.000 claims description 3
- 230000007246 mechanism Effects 0.000 claims description 3
- 238000012216 screening Methods 0.000 claims description 3
- 201000011510 cancer Diseases 0.000 abstract description 12
- 238000000034 method Methods 0.000 abstract description 5
- 238000012864 cross contamination Methods 0.000 abstract description 4
- 230000015572 biosynthetic process Effects 0.000 abstract description 3
- 238000011161 development Methods 0.000 abstract description 3
- 230000008569 process Effects 0.000 abstract description 3
- 238000007877 drug screening Methods 0.000 abstract description 2
- 210000004027 cell Anatomy 0.000 description 104
- 206010061534 Oesophageal squamous cell carcinoma Diseases 0.000 description 8
- 208000036765 Squamous cell carcinoma of the esophagus Diseases 0.000 description 8
- 210000001519 tissue Anatomy 0.000 description 8
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- 239000006285 cell suspension Substances 0.000 description 6
- 230000012010 growth Effects 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 230000010261 cell growth Effects 0.000 description 5
- 208000024891 symptom Diseases 0.000 description 5
- 238000012360 testing method Methods 0.000 description 5
- 108700028369 Alleles Proteins 0.000 description 4
- 239000002054 inoculum Substances 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 230000009747 swallowing Effects 0.000 description 4
- 108020004414 DNA Proteins 0.000 description 3
- 101000581981 Homo sapiens Neural cell adhesion molecule 1 Proteins 0.000 description 3
- 241001465754 Metazoa Species 0.000 description 3
- 102100027347 Neural cell adhesion molecule 1 Human genes 0.000 description 3
- 210000000349 chromosome Anatomy 0.000 description 3
- 230000018044 dehydration Effects 0.000 description 3
- 238000006297 dehydration reaction Methods 0.000 description 3
- 210000003238 esophagus Anatomy 0.000 description 3
- 210000002950 fibroblast Anatomy 0.000 description 3
- 238000011010 flushing procedure Methods 0.000 description 3
- 210000003296 saliva Anatomy 0.000 description 3
- WZUVPPKBWHMQCE-XJKSGUPXSA-N (+)-haematoxylin Chemical compound C12=CC(O)=C(O)C=C2C[C@]2(O)[C@H]1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-XJKSGUPXSA-N 0.000 description 2
- IAKHMKGGTNLKSZ-INIZCTEOSA-N (S)-colchicine Chemical compound C1([C@@H](NC(C)=O)CC2)=CC(=O)C(OC)=CC=C1C1=C2C=C(OC)C(OC)=C1OC IAKHMKGGTNLKSZ-INIZCTEOSA-N 0.000 description 2
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 2
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- WZUVPPKBWHMQCE-UHFFFAOYSA-N Haematoxylin Natural products C12=CC(O)=C(O)C=C2CC2(O)C1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-UHFFFAOYSA-N 0.000 description 2
- 108091092878 Microsatellite Proteins 0.000 description 2
- 241000699660 Mus musculus Species 0.000 description 2
- 241000283973 Oryctolagus cuniculus Species 0.000 description 2
- 208000033240 Progressive symmetric erythrokeratodermia Diseases 0.000 description 2
- 230000001464 adherent effect Effects 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 238000010171 animal model Methods 0.000 description 2
- 230000003915 cell function Effects 0.000 description 2
- 230000004663 cell proliferation Effects 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000002512 chemotherapy Methods 0.000 description 2
- 238000010367 cloning Methods 0.000 description 2
- 230000037213 diet Effects 0.000 description 2
- 235000005911 diet Nutrition 0.000 description 2
- 230000029087 digestion Effects 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 229940000406 drug candidate Drugs 0.000 description 2
- 210000003237 epithelioid cell Anatomy 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 239000012535 impurity Substances 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
- 210000004185 liver Anatomy 0.000 description 2
- 239000003147 molecular marker Substances 0.000 description 2
- 230000003287 optical effect Effects 0.000 description 2
- 230000001575 pathological effect Effects 0.000 description 2
- 210000003800 pharynx Anatomy 0.000 description 2
- 230000002980 postoperative effect Effects 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 238000005070 sampling Methods 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 206010041823 squamous cell carcinoma Diseases 0.000 description 2
- 210000003437 trachea Anatomy 0.000 description 2
- 210000004881 tumor cell Anatomy 0.000 description 2
- 230000005740 tumor formation Effects 0.000 description 2
- 208000008035 Back Pain Diseases 0.000 description 1
- 208000007026 Bronchial fistula Diseases 0.000 description 1
- 206010008479 Chest Pain Diseases 0.000 description 1
- 206010011224 Cough Diseases 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- 206010013952 Dysphonia Diseases 0.000 description 1
- 208000010473 Hoarseness Diseases 0.000 description 1
- 208000016495 Horner Syndrome Diseases 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 206010023126 Jaundice Diseases 0.000 description 1
- 206010027336 Menstruation delayed Diseases 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 206010030113 Oedema Diseases 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 208000002193 Pain Diseases 0.000 description 1
- 102000003992 Peroxidases Human genes 0.000 description 1
- 208000010513 Stupor Diseases 0.000 description 1
- 206010065787 Tracheal fistula Diseases 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 210000000683 abdominal cavity Anatomy 0.000 description 1
- 229960000583 acetic acid Drugs 0.000 description 1
- 238000001467 acupuncture Methods 0.000 description 1
- 208000009956 adenocarcinoma Diseases 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 208000019804 backache Diseases 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 210000000621 bronchi Anatomy 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 230000001612 cachectic effect Effects 0.000 description 1
- 239000012930 cell culture fluid Substances 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 229960001338 colchicine Drugs 0.000 description 1
- 230000005757 colony formation Effects 0.000 description 1
- 230000000093 cytochemical effect Effects 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000009509 drug development Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 230000004743 esophageal carcinogenesis Effects 0.000 description 1
- 239000003172 expectorant agent Substances 0.000 description 1
- 230000003419 expectorant effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 239000000834 fixative Substances 0.000 description 1
- 239000000989 food dye Substances 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 239000012362 glacial acetic acid Substances 0.000 description 1
- 238000002649 immunization Methods 0.000 description 1
- 230000003053 immunization Effects 0.000 description 1
- 238000002991 immunohistochemical analysis Methods 0.000 description 1
- 230000002055 immunohistochemical effect Effects 0.000 description 1
- 238000011532 immunohistochemical staining Methods 0.000 description 1
- 238000003364 immunohistochemistry Methods 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 230000001788 irregular Effects 0.000 description 1
- 238000009940 knitting Methods 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 206010025482 malaise Diseases 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 238000000386 microscopy Methods 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 108040007629 peroxidase activity proteins Proteins 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000001959 radiotherapy Methods 0.000 description 1
- 210000002416 recurrent laryngeal nerve Anatomy 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 210000002345 respiratory system Anatomy 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 230000035807 sensation Effects 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 210000001562 sternum Anatomy 0.000 description 1
- 239000004575 stone Substances 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 238000011477 surgical intervention Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 210000000331 sympathetic ganglia Anatomy 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 239000011534 wash buffer Substances 0.000 description 1
- 230000003442 weekly effect Effects 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0693—Tumour cells; Cancer cells
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/027—New or modified breeds of vertebrates
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
- G01N33/5011—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing antineoplastic activity
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
- G01N33/5044—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics involving specific cell types
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2227/00—Animals characterised by species
- A01K2227/10—Mammal
- A01K2227/105—Murine
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2267/00—Animals characterised by purpose
- A01K2267/03—Animal model, e.g. for test or diseases
- A01K2267/0331—Animal model for proliferative diseases
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2503/00—Use of cells in diagnostics
- C12N2503/02—Drug screening
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Immunology (AREA)
- Chemical & Material Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- Hematology (AREA)
- Molecular Biology (AREA)
- Urology & Nephrology (AREA)
- Cell Biology (AREA)
- Zoology (AREA)
- Biochemistry (AREA)
- Microbiology (AREA)
- General Health & Medical Sciences (AREA)
- Tropical Medicine & Parasitology (AREA)
- Genetics & Genomics (AREA)
- Analytical Chemistry (AREA)
- Medicinal Chemistry (AREA)
- Food Science & Technology (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Organic Chemistry (AREA)
- Toxicology (AREA)
- Wood Science & Technology (AREA)
- Physics & Mathematics (AREA)
- Environmental Sciences (AREA)
- Oncology (AREA)
- Biodiversity & Conservation Biology (AREA)
- Animal Husbandry (AREA)
- General Engineering & Computer Science (AREA)
- Animal Behavior & Ethology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses an esophageal cancer cell line and application thereof. The esophageal cancer cell line is named as a human esophageal squamous cancer cell line ZEC-056 and collected under CCTCC NO: C201691. The human esophageal squamous cancer cell line ZEC-056 comes from tumor tissue of a Chinese patient with esophageal cancer after excision, STR detection results show that the human esophageal squamous cancer cell line ZEC-056 is unique, no cross contamination with other cells occurs in the process of primary culture, clone formation and tumorigenicity is high, and the esophageal cancer cell line can serve as an ideal cell line in the application aspects of esophageal cancer studying, esophageal cancer detection kit development and drug screening.
Description
Technical field
The present invention relates to biology and oncology's technical field, particularly relate to a kind of esophageal carcinoma cell line and application thereof.
Background technology
The esophageal carcinoma is common malignant tumor of digestive tract.The whole world there are about 300,000 people every year and dies from the esophageal carcinoma.Its sickness rate and death
Rate various countries are widely different.China is one of Esophageal Cancer area in the world, and every annual is died of illness about 150,000 people.Man is more than female,
Age of onset is many more than 40 years old.The typical symptom of the esophageal carcinoma is Progressive symmetric erythrokeratodermia acataposis, before this food of difficult dry pharynx, is then
Semiliquid diet, last water and saliva can not be swallowed.In China, Incidence of esophageal cancer occupies general tumour the 5th, and it is dead
Rate occupies the 4th.Two kinds of Main Subtypes of the esophageal carcinoma: scale cancer and adenocarcinoma, wherein scale cancer accounts for 90%.
Patient with esophageal carcinoma clinical manifestation is as follows:
In early days: symptom is the most inconspicuous, discomfort in various degree but may be had to feel, including swallowing food when swallowing thick and stiff food
Choking feeling, burns sample, acupuncture sample or tractive friction sample pain after breastbone.Food is by slow, and has stagnation sense or foreign body sensation.
Stalk is choked to stagnate to feel and is disappeared usually through alleviating after swallowing water.During symptom light time weight, make slow progress.
Middle and advanced stage: the typical symptom of the esophageal carcinoma is Progressive symmetric erythrokeratodermia acataposis, the food of difficult dry pharynx, was semiliquid diet then before this,
Last water and saliva can not be swallowed.Often tell mucoid expectorant, for the saliva swallowed and the secretions of esophagus.Patient gradually becomes thin,
Dehydration, unable.Continuing chest pain or backache is expressed as symptom in late period, cancer has been invaded and has been organized outside esophagus.When cancerous protuberance is blocked caused
Inflammation edema transient remission, or after part cancerous protuberance comes off, obstruction can temporarily alleviate, and constant error thinks that sb.'s illness took a favorable turn.If cancerous protuberance
Invade recurrent laryngeal nerve, may occur in which hoarseness；If compressing cervical sympathetic ganglia, Horner syndrome can be produced；If intrusion trachea,
Bronchus, can form esophagus, trachea or bronchial fistula, acutely chokes and coughs, and respiratory system sense occurs when there is swallowing water or food
Dye.Cachectic states finally occurs.If there being the Organ relative weight such as liver, brain, may occur in which the states such as jaundice, seroperitoneum, stupor.
Treatment main point of surgical intervention, radiotherapy, chemotherapy etc., in recent years, reached based on the Comprehensive Treatment of operation
Arrive bottleneck, and Chemotherapy in Esophageal Cancer slower development, there is no clear and definite conclusion and standard scheme.
For understanding the pathogeny of esophageal squamous cell carcinoma and medicine and the research of other Therapeutic Method in depth, it is necessary to set up suitable diagnostic cast
Type.The most conventional esophageal cancer cell strain is mainly derived from Japan, including KYSE series and TE series.These are thin
Born of the same parents are to pass in vitro for a long time, and some have lost the characteristic of primary tumor tissues.Because race difference, living habit is different,
The reasons such as environmental factors, these cells containing sequences represent the type of China's esophageal carcinoma and the query that feature existence is the biggest.Domestic minority is real
Testing the cell strain set up room less, the widely used cell strain of part is polluted by other cells so that the research of the esophageal carcinoma
Can not carry out in a deep going way.And, along with the accurate medical science of tumor treatment concept go deep into clinical practice, it is provided that the more esophageal carcinoma is thin
Born of the same parents' strain contributes to research and development and the application of targeted drug.Set up the esophageal cancer cell strain in Chinese source, for the morbidity machine of esophageal squamous cell carcinoma
System and treatment provide new model, have important theory and actual application and are worth.
Summary of the invention
The present invention is directed to the current domestic esophageal cancer cell strain lacking Chinese source, and provide one and derive from Chinese food
Pipe cancerous cell line.
From an example patient with esophageal carcinoma, (patient is 63 years old male to the present inventor's esophageal squamous cell carcinoma cell line ZEC-056, and tumor is positioned at food
Pipe stage casing, postoperative pathological breaks up squamous cell carcinoma, TNM T4aN0M0 by stages, phase clinical stages 3A in showing.) operation
Tumor tissues sampling after excision, through original cuiture and after building and being tied to form merit, named Human esophageal squamous cell cancer cell line ZEC-056, in
On May 11st, 2016 is preserved in the China typical culture collection center being positioned at Wuhan, China Wuhan University, and preserving number is:
CCTCC NO:C201691.
The present inventor's esophageal squamous cell carcinoma cell line ZEC-056 cell growth vigorous, clear background, impurity is rare, cell be flat not
Regular polygon, cell is the most closely coupled, meets the feature of epithelioid cell.After adherent, growth is very fast, has good body
Outer cultivation amplification property.Chromosome number is distributed between 38～49 mostly, and mode is 44, meets the feature of malignant tumor.
The cells such as the STR sequencing result of the present inventor's esophageal squamous cell carcinoma cell line ZEC-056 and ATCC, DSMZ preserve the number in storehouse
Carry out inquiry contrast according to storehouse, do not find identical STR testing result, it was demonstrated that it is unique, and does not sends out during original cuiture
The raw cross-contamination with other cells.Immunohistochemical experiment finds, Human esophageal squamous cell cancer cell line ZEC-056 cell is CK5/6 sun
Property, CK14 is positive, and p63 is positive, and CgA is negative, and Sy is negative and CD56 is negative.
Colony formation finds, the cloning efficiency of Human esophageal squamous cell cancer cell line ZEC-056 cell and inoculum density have relation,
Inoculum density is that Cell clonality during 4500 cells is stronger.
Nude mice (nude mouse) becomes tumor experimental result to find, Human esophageal squamous cell cancer cell line ZEC-056 Tumor formation is stronger.
Invention further provides the daughter cell of described esophageal carcinoma cell line.Described daughter cell remains parental generation substantially or all
The characteristic of cell.
Invention further provides described esophageal cancer cell and tie up to the application in the cell model as esophageal carcinoma genesis mechanism research.
Owing to the present inventor's esophageal squamous cell carcinoma cell line ZEC-056 is originated foundation from Chinese, and to build be that the time is shorter, and character is stable,
Using this esophageal carcinoma cell line as study model, have very great help for understanding Chinese primary esophageal carcinogenesis mechanism.
Present invention also offers described esophageal cancer cell to tie up to set up the application in mammal esophageal carcinoma model.Described suckling
Animal is nude mouse or nude rat.By the described Human esophageal squamous cell cancer cell line ZEC-056 cell of certain cell quantity is inoculated in
The positions such as subcutaneous, the liver of nude mouse or nude rat, abdominal cavity or tail vein, it is thus achieved that the animal model of the esophageal carcinoma.
Present invention also offers described esophageal cancer cell to tie up to extract the application in esophageal carcinoma specific tumour mark.By with
Normal esophageal cells and other kinds of cancer cell compare research, it appeared that the molecular marker of the esophageal carcinoma, for this
Molecular marker can carry out the application of the aspects such as follow-up disease detection and drug development.
Present invention also offers described esophageal cancer cell and tie up to the application in screening or assessment treatment esophageal carcinoma medicine.First, logical
Crossing interpolation different pharmaceutical in described Human esophageal squamous cell cancer cell line ZEC-056 culture medium, observation of cell state changes, it is thus achieved that preliminary
Effective drug candidate.Then, drug candidate is administered to the animal model of the above-mentioned esophageal carcinoma, observes and non-dispenser treated animal
Survival period, tumor size, transfer case etc., screening obtains the medicine of the potential treatment esophageal carcinoma.
Present invention also offers described esophageal cancer cell to tie up to develop the application in esophageal carcinoma detection kit.The discovery esophageal carcinoma is special
After the tumor markers of the opposite sex, the detection esophageal carcinoma can be developed according to this tumor markers and occur or the detection kit of development.
Tumor group after the excision of the patient with esophageal carcinoma that the present inventor's esophageal squamous cell carcinoma cell line ZEC-056 comes from an example China
Knitting, STR testing result proves that it is unique, and the cross-contamination with other cells does not occurs during original cuiture, gram
Grand formation is strong with Tumor formation, can be as aspect application such as esophageal carcinoma research and the exploitation of esophageal carcinoma detection kit, drug screenings
Ideal cell line.
Accompanying drawing explanation
Fig. 1 is the Morphology observation figure of Human esophageal squamous cell cancer cell line ZEC-056 cell；
Fig. 2 is the cell growth curve figure of Human esophageal squamous cell cancer cell line ZEC-056；
Fig. 3 is the karyotype scattergram of Human esophageal squamous cell cancer cell line ZEC-056 cell；
Fig. 4 is STR analysis result figure, and wherein, figure A～F is the most homoallelic result figure respectively；
Fig. 5 is Human esophageal squamous cell cancer cell line ZEC-056 cellular immunization CYTOCHEMICAL ANALYSIS result figure；
Fig. 6 is the immunohistochemical analysis result figure of the tumor tissues in source identical with Human esophageal squamous cell cancer cell line ZEC-056；
Fig. 7 is Human esophageal squamous cell cancer cell line ZEC-056 cell clonal formation experimental result picture, wherein, and figure A, B and C inoculation
Cell number is respectively 500,1500 and 4500；
Fig. 8 is that mice becomes tumor experimental result picture.
Detailed description of the invention
Embodiment 1
From an example patient with esophageal carcinoma, (patient is 63 years old male, and tumor is positioned at Esophageal Middle Segment, and it is thin that postoperative pathological breaks up squamous in showing
Born of the same parents' cancer, TNM T4aN0M0 by stages, phase clinical stages 3A.) excision after tumor tissues sampling.Separator well new
Fresh human esophageal carcinoma, after cleaning with PBS, then cuts in culture dish with apparatuses such as ophthalmology tweezers, shears in aseptic super-clean bench
Broken it is separated into 0.5～1mm3Fritter be laid at the bottom of ware, and (GIBCO is public to add the 10%FBS containing 10mL in culture dish
Department), 1% dual anti-DMEM/F12 cell culture medium (GIBCO company), put into 37 DEG C, 5%CO2Incubator in carry out
Cultivate.Change liquid after 5～7 days, discard the downright bad floating fritter come off in piece of tissue, carry out Secondary Culture.In Secondary Culture process
In, utilize fibroblast different from tumor cell digestion power, constantly remove fibroblast.After repeatedly passing on, cultivate
In ware, naked eyes cannot observe fibroblast, and can pass on by continued propagation.Building after being tied to form merit, named Human esophageal squamous cell cancer is thin
Born of the same parents system ZEC-056, was preserved in the China typical culture collection being positioned at Wuhan, China Wuhan University on May 11st, 2016
The heart, preserving number is: CCTCC NO:C201691.
Embodiment 2
Take the Human esophageal squamous cell cancer cell line ZEC-056 cell of Secondary Culture, under an optical microscope (Japan Olympus IMT-2
Inverted microscope) observe living cell growth situation.As it is shown in figure 1, cell Optical Morphology picture, it is seen that cell growth is vigorous,
Clear background, impurity is rare, and cell is flat irregular polygon, and cell is the most closely coupled, meets the spy of epithelioid cell
Point.
Embodiment 3
Cell index (Cell Index) refers to that living cells interacts with microelectrode in detection plate hole, produces the change of electrical impedance,
XCELLigence cell function analyser, these signals are converted into specific parameter becomes cell index.Cell index is fine
Weighed the state growth of cell, diffusion, alteration of form, death, stress wait, cell index is by many magazines
Comment on and adopt.
Take the Human esophageal squamous cell cancer cell line ZEC-056 cell that growth conditions is good, through trypsinization, make cell suspension, and count
Number.The E-Plate detection plate of xCELLigence cell function analyser adds culture medium and measures background impedance value, then exists
E-Plate detection plate adds 100 μ L cell suspension (5000), in room temperature super-clean bench, places 30min.Cell will be added
E-Plate detection plate puts into (monitor station is placed in advance in incubator) on monitor station, carries out the cell proliferation detection of Real-time and Dynamic i.e.
Cell proliferation curve, keeping count 4～6 days can be obtained.Utilize Graph Pad Software on Drawing growth curve, and be calculated thin
The population doubling time of born of the same parents about 16 hours.As in figure 2 it is shown, growth after the present inventor's esophageal squamous cell carcinoma cell line ZEC-056 is adherent
Comparatively fast, there is good cultured and amplified in vitro.
Embodiment 4
Take Human esophageal squamous cell cancer cell line ZEC-056 cell, be planted in six orifice plates, after cultivating 48 hours, use 0.01mg/mL
Colchicine process 16h, when microscopy observes that M phase cell proportion is more than 50%, collect M phase cell.Use KCl
Hypotonic medium Hypotonic treatment M phase cell 15～20min, at room temperature solid as fixative with methanol/glacial acetic acid (volume ratio 3: 1)
Determine cell, film-making on slide.Slide being placed in 0.02% trypsin solution digestion 30～60s, rinses through PBS, Giemsa contaminates
Color, dries, and completes film-making.In basis of microscopic observation, calculate chromosome number in each cell, randomly select 20 cells and enter
Row calculates.As it is shown on figure 3, chromosome number is distributed between 38～49 mostly, mode is 44, meets the feature of malignant tumor.
Embodiment 5
STR (short tandem repeat, STR) is also called microsatellite DNA.General by one long 2～6bp
Core sequence through repeatedly tandem sequence repeats arrangement form, number of repetition is mostly between 10～60 times.The weight of core sequence between individuality
Again count in variability, thus the number of repetition of one group of STR sequence is the most unique in Different Individual, is that cell is raw
The main method that cell identity and source are identified by thing.Human esophageal squamous cell cancer cell line ZEC-056 collecting fresh cultured is thin
Born of the same parents, with Qiagen genomic DNA Mini Kit (purchased from Qiagen company, name of article QIAamp DNA Mini Kit,
Article No. is 51304) extract cell genomic dna, carry out PCR amplification, to gained with 5' end fluorescently-labeled STR primer
Product checks order.Wherein the primer sequence of STR bit point and copy number are as shown in table 1 and Fig. 4.Above-mentioned sequence and ATCC,
The cells such as DSMZ preserve the data base in storehouse and carry out inquiry contrast, do not find identical STR testing result, thus may certify that it is
Uniquely, and the cross-contamination with other cells does not occurs during original cuiture.
The copy number in table 1STR site.
| Labelling (Marker) | Allele 1 (Allele 1) | Allele 2 (Allele 2) |
| TH01 | 9 | 9 |
| D12S391 | 18 | 18 |
| D7S820 | 11 | 13 |
| CSF1PO | 10 | 10 |
| FGA | 22 | 23 |
| AMEL | X | Y |
| D5S818 | 9 | 9 |
| D2S1338 | 22 | 22 |
| D21S11 | 29 | 29 |
| D18S51 | 17 | 17 |
| TPOX | 8 | 8 |
| VWA | 14 | 15 |
| D8S1179 | 10 | 11 |
| D3S1358 | 15 | 17 |
| D13S317 | 8 | 8 |
| D6S1043 | 11 | 14 |
| D16S539 | 9 | 13 |
| PENTAE | 12 | 22 |
| D19S433 | 13 | 15.2 |
| PENTAD | 9 | 12 |
Embodiment 6
SABC detection Human esophageal squamous cell cancer cell line ZEC-056 cellular labeled proteins.S-P method is used to carry out immunohistochemical staining,
Related kit is purchased from Fuzhou Maixin biotechnology Development Co., Ltd.Antibody used be CK5/6 (article No. MAB-0276,
Foochow steps new company), P63 (article No. ZM-04-06, Beijing company of Zhong Shan Golden Bridge), (article No. ZA-0540, in Beijing for CK14
Company of shirt Golden Bridge), CgA (article No. ZA-0507, Beijing company of Zhong Shan Golden Bridge), Sy (article No. IR660, DAKO company),
CD56 (I article No. R628, DAKO company).
Tumor tissues SABC: tissue (for obtaining the same tumor sample of cell in embodiment 1) wax stone dewaxes also through routine
Aquation, uses 3%H2O2Deionized water soaks 10 minutes, to block endogenous peroxydase, is then put into 0.01M pH6.0
Inflexible rubber acid buffer in, pressure cooker spray vapour half a minute carry out antigen retrieval.Dropping one resists, incubated at room temperature 60 minutes, PBS
Wash buffer 3 times, each 2 minutes.Dropping Polymer Helper (polymeric adjuvants), incubated at room temperature 20 minutes,
PBS flushing 3 times, each 2 minutes.Dropping Polyperoxidase-anti-mouse/rabbit IgG, incubated at room temperature
30 minutes, PBS flushing 3 times, each 2 minutes.Dilution preparation DAB solution, develops the color 5～10 minutes, Jing Xiaguan
Examine.Clear water rinses, and haematoxylin is redyed, conventional dehydration, transparent, mounting.
Tumor cell SABC: cell to be detected (Human esophageal squamous cell cancer cell line ZEC-056 cell) shifts to an earlier date 24 hours creep plates
On sterilization microscope slide, after 24 hours, rinsing 2 times with PBS, cold acetone fixes 15 minutes, is soaked in PBS
Standby.Taking the incubated at room temperature 10 minutes that required slide adds 50 μ L peroxidase blocking solution (reagent A), PBS buffers
Liquid rinses 3 times, each 3 minutes, removes PBS liquid, drips the normal nonimmune animal serum of 50 μ L (reagent B), under room temperature
Hatch 10 minutes；Removing serum, drip 50 μ L mono-and resist, incubated at room temperature 60 minutes, PBS flushes three times, each 3～5
Minute；Dropping two resists, and incubated at room temperature 10 minutes, PBS flushes three times, each 3 minutes.Dropping
Polyperoxidase-anti-mouse/rabbit IgG, incubated at room temperature 30 minutes, PBS flushing 3 times, each 2 points
Clock.Dilution preparation DAB solution, develops the color 5～10 minutes, Microscopic observation.Clear water rinses, and haematoxylin is redyed, conventional dehydration,
Transparent, mounting.
As shown in Figure 5 and Figure 6, Human esophageal squamous cell cancer cell line ZEC-056 cell is that CK5/6 is positive to result, and CK14 is positive,
P63 is positive, and CgA is negative, and Sy is negative and CD56 is negative.The immunohistochemistry results with tissue of cell all illustrates that people eats
Pipe squamous cell carcinoma system ZEC-056 is scale cancer source.
Embodiment 7
Take the Human esophageal squamous cell cancer cell line ZEC-056 cell that growth conditions is good, after trypsinization, make cell suspension, and count
Number.By cell suspension with 500,1500,4500 cell per well are inoculated in 6 orifice plates.6 orifice plates are placed in incubator,
Quiescent culture 2 weeks, changes weekly liquid 2 times.After 2 weeks, discarding cell culture fluid, after rinsing with PBS, absolute methanol fixes 10
Min, then by 0.1% violet staining 10min, washes away dyeing liquor, drying at room temperature, and Taking Pictures recording.As it is shown in fig. 7,
The cloning efficiency of Human esophageal squamous cell cancer cell line ZEC-056 cell and inoculum density have relation, and inoculum density is 4500 cells
Time Cell clonality stronger.
Embodiment 8
Take the Human esophageal squamous cell cancer cell line ZEC-056 cell that growth conditions is good, after trypsinization, make cell suspension, and count
Number.Cell suspension is inoculated into 5 nude mice (nude mouse) oxters, Continuous Observation respectively with 10,000,000 cells.Result of the test
As shown in Figure 8, accessible to lump, increase in time after 2 weeks after inoculation, lump increases.
Claims (8)
1. esophageal carcinoma cell line, it is characterised in that named Human esophageal squamous cell cancer cell line ZEC-056, preserving number is: CCTCC
NO:C201691.
2. the daughter cell of esophageal carcinoma cell line as claimed in claim 1.
3. esophageal cancer cell as claimed in claim 1 ties up to the application in the cell model as esophageal carcinoma genesis mechanism research.
4. esophageal cancer cell as claimed in claim 1 ties up to set up the application in mammal esophageal carcinoma model.
Apply the most as claimed in claim 4, it is characterised in that described mammal is nude mouse or nude rat.
6. esophageal cancer cell as claimed in claim 1 ties up to extract the application in esophageal carcinoma specific tumour mark.
7. the application during esophageal cancer cell as claimed in claim 1 ties up to screening or assessment treatment esophageal carcinoma medicine.
8. esophageal cancer cell as claimed in claim 1 ties up to develop the application in esophageal carcinoma detection kit.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610585742.4A CN106011068A (en) | 2016-07-22 | 2016-07-22 | Esophageal cancer cell line and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610585742.4A CN106011068A (en) | 2016-07-22 | 2016-07-22 | Esophageal cancer cell line and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106011068A true CN106011068A (en) | 2016-10-12 |
Family
ID=57116419
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610585742.4A Pending CN106011068A (en) | 2016-07-22 | 2016-07-22 | Esophageal cancer cell line and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106011068A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111662874A (en) * | 2020-06-18 | 2020-09-15 | 上海市胸科医院 | Chinese esophageal squamous carcinoma cell line and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102766599A (en) * | 2012-07-06 | 2012-11-07 | 中国人民解放军第二军医大学 | Application of esophageal squamous cell carcinoma primary tumor strain CH-H-1 |
| CN102766600A (en) * | 2012-07-06 | 2012-11-07 | 中国人民解放军第二军医大学 | Application of esophageal squamous cell carcinoma primary tumor line CH-H-2 |
-
2016
- 2016-07-22 CN CN201610585742.4A patent/CN106011068A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102766599A (en) * | 2012-07-06 | 2012-11-07 | 中国人民解放军第二军医大学 | Application of esophageal squamous cell carcinoma primary tumor strain CH-H-1 |
| CN102766600A (en) * | 2012-07-06 | 2012-11-07 | 中国人民解放军第二军医大学 | Application of esophageal squamous cell carcinoma primary tumor line CH-H-2 |
Non-Patent Citations (6)
| Title |
|---|
| YING-CHUAN HU等: "Establishment and Characterization of HKESC-1, a New Cancer Cell Line from Human Esophageal Squamous Cell Carcinoma", 《CANCER GENET CYTOGENET》 * |
| 卢善明等: "食管癌细胞系 CSEC的建立及其生物学特性", 《实用肿瘤杂志》 * |
| 周勇安等: "人食管癌细胞系的建立及其转移相关基因PCNA的表达", 《细胞与分子免疫学杂志》 * |
| 王秀琴等: "两个人食管癌细胞系的建立及染色体分析", 《中华肿瘤杂志》 * |
| 肖枫等: "四例食管癌细胞系的分子细胞遗传学研究", 《中华医学遗传学杂志》 * |
| 陈晓等: "原代培养建立哈萨克族食管癌细胞系的研究", 《新疆医科大学学报》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111662874A (en) * | 2020-06-18 | 2020-09-15 | 上海市胸科医院 | Chinese esophageal squamous carcinoma cell line and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CA2620616C (en) | Stem cell fusion model of carcinogenesis | |
| CN105296430B (en) | A kind of human colon cancer cells system DXH-1 and its application | |
| CN111793604B (en) | Ornitinib-resistant human non-small cell lung cancer cell strain H1975/OR and application thereof | |
| CN103451148A (en) | Human normal bronchial epithelial cell and preliminary isolated culture and subculture methods and purposes thereof | |
| CN104031884B (en) | The protein arginine transmethylase 7 application in cancer cell metastasis | |
| CN105969734A (en) | Esophageal cancer cell line and application thereof | |
| CN106011067A (en) | Esophageal cancer cell line and application thereof | |
| CN106011069A (en) | Esophageal cancer cell line and application thereof | |
| CN105255832A (en) | Human bile duct cancer cell line and applications thereof | |
| CN106047815A (en) | Esophageal cancer cell line and application thereof | |
| CN106011068A (en) | Esophageal cancer cell line and application thereof | |
| CN115786262B (en) | Human hilar bile duct cancer cell line CBC3T-1 and application thereof | |
| CN116463291A (en) | Human colorectal cancer primary focus and liver transfer focus pairing organoid cell line and culture method and application thereof | |
| CN106011070A (en) | Esophageal cancer cell line and application thereof | |
| CN108251526A (en) | The application of Suppressor of Cytokine Signaling 2 | |
| CN105462928B (en) | A kind of Chinese oral cavity K-1735 COMM-1 and its method for building up | |
| CN106222142A (en) | A kind of esophageal carcinoma cell line and application thereof | |
| CN106190982A (en) | A kind of esophageal carcinoma cell line and application thereof | |
| CN106190981A (en) | A kind of esophageal carcinoma cell line and application thereof | |
| CN106047814A (en) | Esophageal cancer cell line and application thereof | |
| CN106047816A (en) | Esophageal cancer cell line and application thereof | |
| CN100408676C (en) | Establishing and application of scale cancer cell line of purify species New Zealand mice oral cavaty and palatal surface parts | |
| CN113201494B (en) | Mucous membrane melanoma cell and application thereof | |
| CN114832105B (en) | Application of inhibitor of C7orf50 gene or protein | |
| CN109486767B (en) | Human epithelial ovarian cancer cell SHOV4 and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20161012 |