CN117625538A - Human intrahepatic bile duct cancer cell line ICC-X2 and application thereof - Google Patents
Human intrahepatic bile duct cancer cell line ICC-X2 and application thereof Download PDFInfo
- Publication number
- CN117625538A CN117625538A CN202311595934.XA CN202311595934A CN117625538A CN 117625538 A CN117625538 A CN 117625538A CN 202311595934 A CN202311595934 A CN 202311595934A CN 117625538 A CN117625538 A CN 117625538A
- Authority
- CN
- China
- Prior art keywords
- duct cancer
- bile duct
- icc
- intrahepatic bile
- cell line
- 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
- 208000006990 cholangiocarcinoma Diseases 0.000 title claims abstract description 58
- 201000007450 intrahepatic cholangiocarcinoma Diseases 0.000 title claims abstract description 56
- 208000014899 intrahepatic bile duct cancer Diseases 0.000 title claims abstract description 50
- 206010028980 Neoplasm Diseases 0.000 claims abstract description 43
- 230000007246 mechanism Effects 0.000 claims abstract description 14
- 206010027476 Metastases Diseases 0.000 claims abstract description 10
- 230000009401 metastasis Effects 0.000 claims abstract description 10
- 238000010171 animal model Methods 0.000 claims abstract description 7
- 238000003745 diagnosis Methods 0.000 claims abstract description 7
- 239000003814 drug Substances 0.000 claims abstract description 7
- 230000008595 infiltration Effects 0.000 claims abstract description 6
- 238000001764 infiltration Methods 0.000 claims abstract description 6
- 238000004321 preservation Methods 0.000 claims abstract description 6
- 238000012216 screening Methods 0.000 claims abstract description 4
- 229940079593 drug Drugs 0.000 claims description 5
- 206010059866 Drug resistance Diseases 0.000 claims description 4
- 239000003560 cancer drug Substances 0.000 claims description 2
- 230000001413 cellular effect Effects 0.000 claims 1
- 210000004027 cell Anatomy 0.000 abstract description 103
- 201000011510 cancer Diseases 0.000 abstract description 13
- 238000011161 development Methods 0.000 abstract description 5
- 230000005856 abnormality Effects 0.000 abstract description 3
- 230000004069 differentiation Effects 0.000 abstract description 3
- 210000004102 animal cell Anatomy 0.000 abstract description 2
- 230000000813 microbial effect Effects 0.000 abstract description 2
- 241000699660 Mus musculus Species 0.000 description 8
- 210000000349 chromosome Anatomy 0.000 description 8
- 238000011580 nude mouse model Methods 0.000 description 8
- 230000001575 pathological effect Effects 0.000 description 7
- 210000001519 tissue Anatomy 0.000 description 7
- 238000002474 experimental method Methods 0.000 description 6
- 238000011160 research Methods 0.000 description 6
- 210000004881 tumor cell Anatomy 0.000 description 6
- 238000004458 analytical method Methods 0.000 description 5
- 230000012010 growth Effects 0.000 description 5
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 4
- 210000003445 biliary tract Anatomy 0.000 description 4
- 230000018109 developmental process Effects 0.000 description 4
- 201000010099 disease Diseases 0.000 description 4
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 230000003698 anagen phase Effects 0.000 description 3
- 230000010261 cell growth Effects 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 230000029087 digestion Effects 0.000 description 3
- 239000012634 fragment Substances 0.000 description 3
- 230000002068 genetic effect Effects 0.000 description 3
- 238000001727 in vivo Methods 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 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
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 238000002738 Giemsa staining Methods 0.000 description 2
- 108091092878 Microsatellite Proteins 0.000 description 2
- 239000002246 antineoplastic agent Substances 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 210000000013 bile duct Anatomy 0.000 description 2
- 238000004113 cell culture Methods 0.000 description 2
- 239000006285 cell suspension Substances 0.000 description 2
- YRQNKMKHABXEJZ-UVQQGXFZSA-N chembl176323 Chemical compound C1C[C@]2(C)[C@@]3(C)CC(N=C4C[C@]5(C)CCC6[C@]7(C)CC[C@@H]([C@]7(CC[C@]6(C)[C@@]5(C)CC4=N4)C)CCCCCCCC)=C4C[C@]3(C)CCC2[C@]2(C)CC[C@H](CCCCCCCC)[C@]21C YRQNKMKHABXEJZ-UVQQGXFZSA-N 0.000 description 2
- 229940044683 chemotherapy drug Drugs 0.000 description 2
- 238000012864 cross contamination Methods 0.000 description 2
- 210000004748 cultured cell Anatomy 0.000 description 2
- 210000002919 epithelial cell Anatomy 0.000 description 2
- 230000003203 everyday effect Effects 0.000 description 2
- 229960005277 gemcitabine Drugs 0.000 description 2
- SDUQYLNIPVEERB-QPPQHZFASA-N gemcitabine Chemical compound O=C1N=C(N)C=CN1[C@H]1C(F)(F)[C@H](O)[C@@H](CO)O1 SDUQYLNIPVEERB-QPPQHZFASA-N 0.000 description 2
- 239000012188 paraffin wax Substances 0.000 description 2
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 230000008261 resistance mechanism Effects 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 230000004083 survival effect Effects 0.000 description 2
- 108700028369 Alleles Proteins 0.000 description 1
- 102000007325 Amelogenin Human genes 0.000 description 1
- 108010007570 Amelogenin Proteins 0.000 description 1
- 238000011729 BALB/c nude mouse Methods 0.000 description 1
- 108090000145 Bacillolysin Proteins 0.000 description 1
- 206010004593 Bile duct cancer Diseases 0.000 description 1
- 208000005623 Carcinogenesis Diseases 0.000 description 1
- 201000009030 Carcinoma Diseases 0.000 description 1
- 108020004414 DNA Proteins 0.000 description 1
- 208000005189 Embolism Diseases 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 102000035092 Neutral proteases Human genes 0.000 description 1
- 108091005507 Neutral proteases Proteins 0.000 description 1
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 229930012538 Paclitaxel Natural products 0.000 description 1
- 108010019160 Pancreatin Proteins 0.000 description 1
- 102100023361 SAP domain-containing ribonucleoprotein Human genes 0.000 description 1
- 108010087230 Sincalide Proteins 0.000 description 1
- 101000693530 Staphylococcus aureus Staphylokinase Proteins 0.000 description 1
- 101710139423 THO complex subunit 1 Proteins 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 229960000583 acetic acid Drugs 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 238000004061 bleaching Methods 0.000 description 1
- 230000036952 cancer formation Effects 0.000 description 1
- 231100000504 carcinogenesis Toxicity 0.000 description 1
- 238000010609 cell counting kit-8 assay Methods 0.000 description 1
- 210000003855 cell nucleus Anatomy 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 230000035572 chemosensitivity Effects 0.000 description 1
- 229960001338 colchicine Drugs 0.000 description 1
- 230000030944 contact inhibition Effects 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 239000006059 cover glass Substances 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 201000008819 extrahepatic bile duct carcinoma Diseases 0.000 description 1
- 210000003194 forelimb Anatomy 0.000 description 1
- 230000007614 genetic variation Effects 0.000 description 1
- 239000012362 glacial acetic acid Substances 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 201000007270 liver cancer Diseases 0.000 description 1
- 208000014018 liver neoplasm Diseases 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 239000002547 new drug Substances 0.000 description 1
- 230000024121 nodulation Effects 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000000771 oncological effect Effects 0.000 description 1
- 229960001756 oxaliplatin Drugs 0.000 description 1
- DWAFYCQODLXJNR-BNTLRKBRSA-L oxaliplatin Chemical compound O1C(=O)C(=O)O[Pt]11N[C@@H]2CCCC[C@H]2N1 DWAFYCQODLXJNR-BNTLRKBRSA-L 0.000 description 1
- 229960001592 paclitaxel Drugs 0.000 description 1
- 229940055695 pancreatin Drugs 0.000 description 1
- 230000008506 pathogenesis Effects 0.000 description 1
- 229910052697 platinum Inorganic materials 0.000 description 1
- 238000004393 prognosis Methods 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000003908 quality control method Methods 0.000 description 1
- 238000001959 radiotherapy Methods 0.000 description 1
- 239000012925 reference material Substances 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 239000012679 serum free medium Substances 0.000 description 1
- IZTQOLKUZKXIRV-YRVFCXMDSA-N sincalide Chemical compound C([C@@H](C(=O)N[C@@H](CCSC)C(=O)NCC(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(N)=O)NC(=O)[C@@H](N)CC(O)=O)C1=CC=C(OS(O)(=O)=O)C=C1 IZTQOLKUZKXIRV-YRVFCXMDSA-N 0.000 description 1
- 239000002356 single layer Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000010183 spectrum analysis Methods 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 239000007929 subcutaneous injection Substances 0.000 description 1
- 238000010254 subcutaneous injection Methods 0.000 description 1
- 238000011521 systemic chemotherapy Methods 0.000 description 1
- RCINICONZNJXQF-MZXODVADSA-N taxol Chemical compound O([C@@H]1[C@@]2(C[C@@H](C(C)=C(C2(C)C)[C@H](C([C@]2(C)[C@@H](O)C[C@H]3OC[C@]3([C@H]21)OC(C)=O)=O)OC(=O)C)OC(=O)[C@H](O)[C@@H](NC(=O)C=1C=CC=CC=1)C=1C=CC=CC=1)O)C(=O)C1=CC=CC=C1 RCINICONZNJXQF-MZXODVADSA-N 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 230000005740 tumor formation Effects 0.000 description 1
- 208000010576 undifferentiated carcinoma Diseases 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
- A01K67/0271—Chimeric vertebrates, e.g. comprising exogenous cells
-
- 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
- C12N5/067—Hepatocytes
-
- 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
- G01N33/5067—Liver 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
- A01K2207/00—Modified animals
- A01K2207/12—Animals modified by administration of exogenous 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
- 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
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2500/00—Screening for compounds of potential therapeutic value
- G01N2500/10—Screening for compounds of potential therapeutic value involving cells
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Cell Biology (AREA)
- Biotechnology (AREA)
- Immunology (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Genetics & Genomics (AREA)
- Urology & Nephrology (AREA)
- Organic Chemistry (AREA)
- Hematology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Gastroenterology & Hepatology (AREA)
- Environmental Sciences (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Toxicology (AREA)
- Tropical Medicine & Parasitology (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- General Engineering & Computer Science (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Animal Behavior & Ethology (AREA)
- Oncology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
Human intrahepatic bile duct cancer cell line ICC-X2 and application thereof, belonging to the field of microbial animal cell line. A novel human intrahepatic bile duct cancer cell line is provided, which is named ICC-X2 and has the preservation number: cctccc NO: C202259.ICC-X2 can be used in establishing cell model of intrahepatic duct cancer occurrence, development or metastasis. ICC-X2 can be applied to establishing an animal model of intrahepatic cholangiocarcinoma. ICC-X2 can be applied to a cell model for researching differentiation mechanism, cell morphology and function abnormality, tumor infiltration and metastasis mechanism of intrahepatic cholangiocarcinoma and guiding clinical comprehensive diagnosis and treatment. ICC-X2 can be applied to researching the occurrence mechanism of intrahepatic duct cancer and screening medicaments for preventing and treating intrahepatic duct cancer. ICC-X2 can be used as a cell model for researching differentiation mechanism, cell morphology and function abnormality, tumor infiltration and metastasis mechanism of intrahepatic cholangiocarcinoma, guiding clinical comprehensive diagnosis and treatment and the like.
Description
Technical Field
The invention belongs to the field of microbial animal cell lines, and particularly relates to a human intrahepatic bile duct cancer cell line ICC-X2 and application thereof.
Background
Intrahepatic cholangiocarcinoma (Intrahepatic Cholangiocarcinoma, ICC) is the second most common type of liver cancer, and the incidence of this disease has been rising worldwide. The tumor originates from biliary tract epithelial cells, which account for 10% -20% of all bile duct malignant tumors; the operation has possible cure only in early stage of the disease, and patients (70% -90%) which are unresectable in late stage have poorer prognosis, and the survival time after diagnosis is less than 12 months. Systemic chemotherapy and radiotherapy with gemcitabine alone or in combination with platinum-based chemotherapeutics have limited effectiveness in improving long-term survival in patients.
Human tumor cell lines, particularly those with complete data and follow-up, are important tools in tumor biology research. Since the 80 s of the 20 th century, a plurality of bile duct malignant tumor cell lines were established and identified by three kingdoms of China, japan and Korean, but few cell lines mainly including extrahepatic bile duct cancer and intrahepatic bile duct cancer have been reported.
Cell lines from primary tumors of the biliary tract system have great roles in exploring biology and biliary tract tumorigenesis, detecting drug sensitivity, developing molecular therapeutic targets and researching drug resistance mechanisms. A complete pool of tumor cell lines should reflect the diversity of tumor phenotypes and be able to provide cell lines of different tumor heterogeneity; given the different etiologies of tumors of the biliary system and their genetic variations related to etiology and race, it is important to use an appropriate preclinical model reflecting these characteristics.
In 2002, a first example of intrahepatic bile duct cancer cell line was built in China. Provides an important cell experimental model for basic and clinical research of national intrahepatic cholangiocarcinoma. However, as the number of in vitro passages increases, some of the unique biological characteristics of the cell line gradually change or disappear, and even characteristics that the originating cell does not have. In addition, many cell lines are detected with cross-contamination of cells, the most common sources of contaminating cells being HeLa, T-24 and M14, and erroneous results from studies using these cells. The national experiment cell resource sharing service platform finds that a plurality of tumor cell lines established by domestic scholars are cross-contaminated when carrying out identity authentication in the processes of collecting, arranging, quality control and preserving cells. A total of 46 samples of 37 lines of cells were collected from different areas and different laboratories, and each of the samples was subjected to species identification and STR spectrum analysis. The cell cross contamination phenomenon is found to be serious, and the error rate reaches 62.6%. And many thousands of papers based on these contaminated cells are in our country each year. In the united states, about 560 billions of dollars are wasted each year due to the lack of reproducibility of preclinical studies. The cost of the biological reagents and reference materials is about one third of the total cost. Cell lines that are misidentified, contaminated, genetically-bleaching, and clonally evolved are important reasons for the inability of many studies to replicate and affect everyone engaged in the cell study.
The newly established cell line can maintain the characteristics similar to those of the primary tumor to the greatest extent, and the result obtained by using the cell line for research is closest to the actual condition of a human body. Because of different genetic backgrounds, it is necessary to build a disease model specific to Chinese in order to better study the disease of Chinese. For the above reasons, the continuous establishment of new cell lines and the elimination of old cell lines have become an important link in the research of pathogenesis and treatment methods of intrahepatic cholangiocarcinoma.
Disclosure of Invention
The invention aims to provide a novel human intrahepatic bile duct cancer cell line ICC-X2 and application thereof, aiming at the problem of insufficient traditional Chinese-source intrahepatic bile duct cancer cell line.
The invention provides a novel human intrahepatic bile duct cancer cell line, which is human intrahepatic bile duct cancer cell line ICC-X2 with the preservation number: cctccc NO: C202259.
the human intrahepatic bile duct cancer cell line ICC-X2 can be applied to establishing a cell model of intrahepatic bile duct cancer occurrence, development or metastasis.
The human intrahepatic bile duct cancer cell line ICC-X2 can be applied to the establishment of an intrahepatic bile duct cancer animal model.
The human intrahepatic bile duct cancer cell line ICC-X2 can be applied to a cell model for researching the differentiation mechanism, cell morphology and function abnormality, tumor infiltration and metastasis mechanism of intrahepatic bile duct cancer and guiding clinical comprehensive diagnosis and treatment.
The human intrahepatic bile duct cancer cell line ICC-X2 can be applied to researching the occurrence mechanism of intrahepatic bile duct cancer and screening medicaments for preventing and treating intrahepatic bile duct cancer.
The invention adopts the operation of removing the primary focus of the specimen from the intrahepatic duct cancer of a 62-year-old female, and uses the mixed digestion of the two collagenase type II and neutral proteinase as primary culture, and establishes a intrahepatic duct cancer cell line named ICC-X2 through cell culture technology. Has been preserved in 22 days of 2 months 2022, the preservation unit: china center for type culture Collection, accession number: university of martial arts, deposit number: cctccc NO: C202259.
the cell line has the following biological characteristics:
1. the cell grows by adherence, is inhibited in a non-contact way, and can generate superposition growth phenomenon.
2. Cell doubling time was approximately 48h.
3. Cell STR results showed: ICC-X2 cells were consistent with STR results from tumor tissue of the same patient, cells were derived from the same tumor sample, and were not contaminated with other cells during culture.
4. Chromosome analysis suggests: the cell of the strain is mainly of a subtetraploid karyotype, and the chromosome number and morphology are greatly different.
5. All the nude mice become tumor after being inoculated with the cells, and the histology of the transplanted tumor is similar to that of the primary tumor.
6. Cell sensitive results to common clinical chemotherapeutic drugs.
The human intrahepatic bile duct cancer cell line ICC-X2 can be used as a cell model for researching occurrence and development of intrahepatic bile duct cancer, tumor infiltration and metastasis mechanisms, tumor drug resistance mechanisms, new drug research and development, guiding clinical comprehensive diagnosis and treatment and the like.
Drawings
FIG. 1 shows the pathological results of ICC-X2 cell-derived tumor tissue.
FIG. 2 is a view of the morphology of ICC-X2 cells under a microscope (. Times.100).
FIG. 3 shows the growth curve of ICC-X2 cells.
FIG. 4 shows the results of ICC-X2 cell chromosome analysis.
FIG. 5 shows the results of an ICC-X2 cell immunodeficient mouse in vivo oncologic assay.
FIG. 6 shows pathological section results (. Times.200) of nude mice transplanted tumors.
FIG. 7 shows the chemosensitivity results of ICC-X2 cells.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention will be further described with reference to the following examples. The experimental methods described in the examples below are all conventional methods without specific description.
The intrahepatic bile duct cancer cell line provided by the invention is a human intrahepatic bile duct cancer cell line ICC-X2, and the preservation number is as follows: cctccc NO: C202259. the primary focus of the specimen is excised by adopting a 62-year-old female intrahepatic duct cancer operation, and is subjected to mixed digestion by two collagenase type II/neutral protease to be subjected to primary culture, and a intrahepatic duct cancer cell line named ICC-X2 is established by a cell culture technology. Has been preserved in 22 days of 2 months 2022, the preservation unit: china center for type culture collection (CCTCC for short, collection address: university of Wuhan, china), accession number: cctccc NO: C202259.
the human intrahepatic bile duct cancer cell line ICC-X2 can be used as a cell model for researching the occurrence, development or metastasis mechanism of intrahepatic bile duct cancer. The human intrahepatic bile duct cancer cell line ICC-X2 can also be used for establishing an intrahepatic bile duct cancer animal model.
1. Establishment of human intrahepatic bile duct cancer cell line ICC-X2
Tumor tissues of patients with intrahepatic duct cancer are clinically collected and subjected to primary culture after being digested by mixed enzymes, so that a strain of intrahepatic duct cancer cell line capable of continuous passage is successfully established, and the cell characteristics are kept stable until the current generation of 102. The pathological result of the patient is medium-low differentiated intrahepatic cholangiocarcinoma, and part of the pathological result is undifferentiated carcinoma. The pathological results of ICC-X2 cell-derived tumor tissue are shown in FIG. 1.
2. Biological property detection of human intrahepatic bile duct cancer cell line ICC-X2
1. Cell morphology: after the cell growth is stable and transferred, living cell observation is carried out on the cultured cells; and fixing the monolayer cells growing on the cover glass with 95% ethanol by volume fraction, staining with HE, and observing with a light microscope. The results show that: cells were found to be arranged as epithelial cells under a phase contrast microscope, and were grown by adherence to the wall with overlapping growth, as shown in FIG. 2.
2. Short fragment repeat (STR) identification
The short tandem repeat sequence is also called microsatellite DNA, and refers to a DNA sequence formed by tandem repeat (the number of times of repeat is 10-60 times, and the gene fragment is below 400 base pairs) on a chromosome, wherein a plurality of base pairs are taken as core units (2-6 base pairs); individual differences in the number of repetitions per core unit can occur, resulting in alleles of differing fragment lengths. Thus, the number of repetitions of a set of STR sequences is almost unique among individuals, and is a genetic identity characteristic of individuals, and is also the primary method of cell biology to identify cell identity and origin.
Fresh cultured ICC-X2 cells and tumor tissue samples of the same patient were collected, genomic DNA was extracted, STR detection service was provided by Souzhou authentication biotechnology Co., ltd, PCR amplification was performed using 5' -end fluorescent-labeled primers, and the resulting products were sequenced, and analyzed for the number of sequence repeats including 20 STR sites such as Amelogenin, THO1, TPOX, D13S317, vWA, D16S539, D5S818, CSF1PO, and D7S 820. The above sequences were compared with databases of cell banks such as ATCC, DSMZ, etc., and the same genetic map was not returned. The ICC-X2 cells were consistent with the STR results of tumor tissue of the same patient, indicating that the cells were derived from the same tumor sample and that ICC-X2 cells were not contaminated with other cells during the culture process, as shown in Table 1.
TABLE 1
3. Cell growth curve assay: taking 18 th generation cells in logarithmic growth phase to obtain 1×10 5 Per ml of single cell suspension, then inoculated in 96-well plates at 100. Mu.L/well, 3 multiplex wells were set. After 24 hours, the OD value at 490nm wavelength was measured by starting the dosing, and the solution was changed every day for 9 days. The growth curve is plotted after the end of the experiment and the doubling time is calculated according to the formula td=t×lg2/lg (N1/N0) as shown in fig. 3. The results show that: the growth of the cells is slow on the 1 st day after adherence, the cells start to grow exponentially on the 2 nd day, and the cells can grow in a superposition way under the condition of sufficient nutrition when the culture medium is replaced every day, and the phenomenon of non-contact inhibition is avoided; the average doubling time of the cells was about 48h.
4. Chromosome analysis: the 30 th generation cells in logarithmic growth phase were taken for karyotyping. Cells were subjected to colchicine for 2h, hypotonic with 0.075mol/L KCl, methanol-glacial acetic acid fixation, ice-wet sheet dripping, room temperature aging, pancreatin treatment, giemsa staining and banding analysis, and the results are shown in FIG. 4, and can be seen from FIG. 4: through Giemsa staining banding chromosome karyotype analysis, the cell line chromosome subtetraploid karyotype is mainly, the chromosome number and morphology are greatly different, and the cell line has the characteristics of malignant tumor cells.
The 4 points above show that the human intrahepatic bile duct cancer cell line ICC-X2 can be applied to a cell model for intrahepatic bile duct cancer occurrence, development or metastasis.
5. In vivo nodulation experiments in immunodeficient mice: 3 female BALB/C nude mice of 4 weeks of age were taken,subcutaneous injections of 1X 10 into the right forelimb 7 Mu.l of cultured cells were used, nude mice were sacrificed by cervical dislocation after 4 weeks, transplanted tumors were removed, formalin fixed, paraffin embedded sections were cut, HE stained, observed under a mirror, and experimental data were recorded by photographing. The results show that: 1 week after subcutaneous inoculation of cells in nude mice, the transplanted tumors can grow out, and all 3 nude mice become tumors, as shown in fig. 5, which shows that the cell line has high tumor formation capability, and the human intrahepatic bile duct cancer cell line ICC-X2 can be applied to establishment of intrahepatic bile duct cancer animal models.
6. Pathological section of tumor: tumor tissue was routinely fixed in formalin, paraffin embedded, sectioned and HE stained. The pathological section results of nude mice transplanted tumor are shown in fig. 6. The results show that: the nude mice are observed under the tumor specimen lens in vivo to have vigorous cell growth, enlarged cell nucleus, deep dyeing and obvious atypical, and the histological morphology of the nude mice is similar to that of clinical specimens. And simultaneously, the phenomena of tumor cell cancer embolism, nerve invasion, satellite range and the like can be seen under the lens. Experiments show that the human intrahepatic bile duct cancer cell line ICC-X2 can be applied to research on tumor infiltration transfer mechanisms of intrahepatic bile duct cancers and animal models for guiding clinical comprehensive diagnosis and treatment.
7. Clinical chemotherapy drug sensitivity detection: ICC-X2 cells in the logarithmic growth phase are collected and are prepared into single cell suspension after trypsin digestion. 10000 cells per well per 100 μl were seeded into 96-well plates and 6 wells per group were replicated. After the cells adhere to the wall, the anti-tumor drugs with different concentrations are added into the experimental group, and the corresponding equal volume of drug dissolution solution is added into the control group. After 72h of drug action, the complete medium was replaced with 100. Mu.l of serum-free medium containing 10% (v/v) CCK-8. After 2h, the OD at 450nm was measured. ICC-X2 cells, as shown in FIG. 7, were sensitive to oxaliplatin (IC50=2.9. Mu. Mol/L) (panel D in FIG. 7) and paclitaxel (IC50=7.3 ng/ml) (panel A in FIG. 7), and were resistant to gemcitabine (IC50 > 60. Mu. Mol/L) (panel C in FIG. 7) and 5-FU (IC50=176.9. Mu. Mol/L) (panel B in FIG. 7). Experiments show that the human intrahepatic bile duct cancer cell line ICC-X2 can be applied to research on the occurrence mechanism of intrahepatic bile duct cancer drug resistance and screening of medicines for preventing and treating intrahepatic bile duct cancer.
The above-described embodiments are merely preferred embodiments of the present invention and should not be construed as limiting the scope of the present invention. All equivalent changes and modifications within the scope of the present invention are intended to be covered by the present invention.
Claims (5)
1. The human intrahepatic bile duct cancer cell line ICC-X2 is characterized in that the human intrahepatic bile duct cancer cell line ICC-X2 is preserved in China center for type culture collection, and the preservation number is CCTCC NO: C202259.
2. use of the human intrahepatic bile duct cancer cell line ICC-X2 according to claim 1 for the establishment of a cellular model of intrahepatic bile duct cancer occurrence, progression or metastasis.
3. Use of the human intrahepatic bile duct cancer cell line ICC-X2 according to claim 1 for the establishment of an intrahepatic bile duct cancer animal model.
4. The use of human intrahepatic bile duct cancer cell line ICC-X2 according to claim 1 for studying tumor infiltration metastasis mechanism of intrahepatic bile duct cancer and guiding animal model of clinical comprehensive diagnosis and treatment.
5. The use of the human intrahepatic bile duct cancer cell line ICC-X2 as claimed in claim 1 for researching the occurrence mechanism of intrahepatic bile duct cancer drug resistance and screening medicines for preventing and treating intrahepatic bile duct cancer.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311595934.XA CN117625538A (en) | 2023-11-28 | 2023-11-28 | Human intrahepatic bile duct cancer cell line ICC-X2 and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311595934.XA CN117625538A (en) | 2023-11-28 | 2023-11-28 | Human intrahepatic bile duct cancer cell line ICC-X2 and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN117625538A true CN117625538A (en) | 2024-03-01 |
Family
ID=90037072
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202311595934.XA Pending CN117625538A (en) | 2023-11-28 | 2023-11-28 | Human intrahepatic bile duct cancer cell line ICC-X2 and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117625538A (en) |
-
2023
- 2023-11-28 CN CN202311595934.XA patent/CN117625538A/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN114717191B (en) | Human intrahepatic bile duct cancer cell strain ICC-X3 and application thereof | |
| Ouyang et al. | Immortal human pancreatic duct epithelial cell lines with near normal genotype and phenotype | |
| Wistuba et al. | Comparison of features of human breast cancer cell lines and their corresponding tumors. | |
| CN114606194B (en) | Human intrahepatic bile duct cancer cell strain ICC-X1 and application thereof | |
| Hu et al. | Establishment, characterization, karyotyping, and comparative genomic hybridization analysis of HKESC-2 and HKESC-3: two newly established human esophageal squamous cell carcinoma cell lines | |
| CN105255832B (en) | Human bile duct cancer cell line and application thereof | |
| CN107541494B (en) | Human bile duct cancer cell line and application thereof | |
| CN110106150B (en) | Preparation method and application of synovial sarcoma cell line hSS-005R | |
| CN117625538A (en) | Human intrahepatic bile duct cancer cell line ICC-X2 and application thereof | |
| CN115786262B (en) | Human hilar bile duct cancer cell line CBC3T-1 and application thereof | |
| Kawashima et al. | Establishment and characterization of a novel myxofibrosarcoma cell line | |
| CN107460171B (en) | Human thyroid undifferentiated cancer cell line and application thereof | |
| CN117736993A (en) | Human pancreatic cancer cell line PDAC-X3 and application thereof | |
| WO2008093886A1 (en) | Mcf7-derived cell | |
| CN111004782B (en) | Primary human intestinal cancer cell and culture method and application thereof | |
| CN104403996B (en) | Human gastric cancer cell line with 5-fluorouracil resistance and establishment method and application thereof | |
| CN110904043A (en) | Primary cells of human intestinal cancer, application and culture method | |
| CN107541495B (en) | FGF19 over-expressed human liver cancer cell line and application thereof | |
| CN117165534B (en) | Immortalized human gastric cancer fibroblast strain and construction and proliferation strengthening method thereof | |
| CN104403997B (en) | Human gastric cancer cell line with cisplatin resistance and establishment method and application thereof | |
| CN114959041B (en) | Novel target and diagnostic marker for inhibiting colorectal cancer proliferation metastasis and application thereof | |
| CN117660656B (en) | Microbial community marker related to prognosis of gastric cancer and application thereof | |
| CN112831472B (en) | Primary human thymoma cell, culture method and application thereof, and serum-free culture medium | |
| CN113430170B (en) | Primary human intestinal cancer cell and culture method and application thereof | |
| CN110835623B (en) | Primary platinum-resistant human ovarian cancer cell line FDOVL, and preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |