CN103305467B - A kind of human liver cancer cell system HLCZ02 and application thereof - Google Patents
A kind of human liver cancer cell system HLCZ02 and application thereof Download PDFInfo
- Publication number
- CN103305467B CN103305467B CN201310214644.6A CN201310214644A CN103305467B CN 103305467 B CN103305467 B CN 103305467B CN 201310214644 A CN201310214644 A CN 201310214644A CN 103305467 B CN103305467 B CN 103305467B
- Authority
- CN
- China
- Prior art keywords
- hlcz02
- liver cancer
- cell system
- cancer cell
- human liver
- 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.)
- Active
Links
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Does the present invention disclose a kind of human liver cancer cell system HLCZ02, and this human liver cancer cell system HLCZ02 is CCTCC for being preserved in China typical culture collection center and preserving number? the human liver cancer cell system of NO:C201310. The human liver cancer cell system HLCZ02 of the present invention can be used as cell model or animal model is applied, and also can be used for preparation, screening or evaluates anti-hepatitis virus medicament, antitumor drug, also can be used for preparing artificial liver.
Description
Technical field
The present invention relates to microorganism zooblast field, particularly relate to a kind of human liver cancer cell system and application thereof.
Background technology
Liver has multiple important physiological function, mainly comprises: synthesizes and secretes a large amount of serum proteins, such as albumin and lipoprotein; The lipid of synthesis and transhipment and protein binding; Function of detoxification; Synthesis secretion bile; Synthesis sugar regulates blood sugar; Decompose amino acid urea synthesis; Activate VITAMIN; Synthesis and decomposition glycogen; Synthesizing glutathion and metallothionein(MT) etc.
Along with the progress of biotechnology, utilize the method for gene recombination and cytogamy can produce much useful biological products, but in contrast, animal cell culture technology seems more important than in the past. Consider that liver has physiological function more more than other organs, comprise the function of its synthesis significantly and secretion plasmosin, and having stronger synthesis albumin, lipoprotein, transport lipid, urea, glycogen, the ability of gsh and stronger metabolic capacity, this makes, and investigator is more interesting to be utilized liver cell to cultivate to produce the said products. Based on this, in order to study liver function and produce relevant biological products, it is necessary to culture hepatocyte. But, current cell culture technology can not produce the biological products such as plasma proteins at the cultivation period of liver cell, and normal liver cell is from the very fast decline of vigor after body, and this causes the acquisition of related products to become very difficult.
Liver is also the organ that in adult body, minority can regenerate, but the liver cell passage number cultivated in culture dish is limited, can not repeatedly cultivate.
More existing hepatic cell lines due to be inhuman source or differ relatively big with human liver cell and can not widespread use. More existing hepatic cell lines deriving from laboratory animal, such as derive from rat (Tsaoet.al., Exp.CellRes., 1984,154:38-52 at present; Enatetal., Proc.Nat.Acad.SciUSA, 1984,87:1411-1415), utilize the rat epithelial cell (ChessebeufandPadieu, InVitro, the 1984,20:780-795 that derive from adult rat liver that serum free medium is set up; Enatetal., Proc.Natl.Acad.Sci., 1984,81:1411-1415). Rat hepatocytes also can proceed to SV40DNA(Woodworthetal., CancerRes., 1987,46:4018-4026; Ledleyetal., Proc.Nat.Acad.Sci.USA, 1987,84:5335-5339), but due to the liver cell metabolism with people different, this kind of cell can not be used in researching human body drug metabolism and liver cancer generates.
In addition, human liver cell vegetative propagation also existing relevant report (KaighnandPrince, Proc.Nat.Acad.Sci., 1971; 68:2396-2400), the people tire liver of long-term cultivation also sets up (Salas-Prato, M.etal.; InVitroCellDev.Biol., 1988,24:230-238; Sells, M.A.etal., InVitroCellDev.Biol., 1985,21:216-220), but due to difference in metabolism of tire liver and adult liver, tire liver is limited to the application in toxicological study in tumour generation, and adult liver is more amenable for use with tumour and generates and toxicological study.
The research model substituting liver cell to find, investigator establishes and derives from liver cancer and have the clone of identical function (such as secreting albumin function) with normal liver cell. multiple hepatoma cell line (e.g., Knowlesetal., U.S.Pat.No.4,393,133, issuedJul.12,1983 are set up at present, KnowlesB.B.etal., Science, 1980,209:497-499, MonjardinoJ.andCrawfordE., Virology, 1979,96:652-655, ParkJ.G.etal., Int.J.Cancer, 1995,62:276-282, Zhongetal., PNAS, 2005,102 (26): 9294-9299, FuandCheng, AntimicrobialAgentsandChemotherapy, 2000,44 (12): 3402-3407), such as hepatoma cell line HepG2(U.S.Pat.No.4393133). utilize research also existing relevant report (Kellyetal., InVitroCell.andDev.Biol., the 1989,25:217-222 of HepG2 further, U.S.Pat.No.5,290,684, andDarlingtonetal., InVitroCell.andDev.Biol., 1987,2-3:349-354). in addition, Nakabayashi establishes hepatoma cell line Huh7(CancerResearch, 1982,42:3858-3863). take a broad view of existing publication document, existing hepatoma cell line includes but not limited to: HLF (OkayamaUniversity, medicalschool:1975), HLE, c-1 (OkayamaUniversity, medicalschool:1975), HuH-6clone5 (OkayamaUniversity, medicalschool:1976), HuH-7 (OkayamaUniversity, medicalschool:1979), C-HC-4 (HokkaidoUniversity, schoolofmedicine:1979), HCC-M (KeioUniversity, schoolofmedicine:1980), JHH-1 (TheTokyoJikeiUniversitySchoolofMedicine:1980), JHH-2 (TheTokyoJikeiUniversitySchoolofMedicine:1982), JHH-4 (TheTokyoJikeiUniversitySchoolofMedicine:1983), KIM-1 (KurumeUniversity, schoolofmedicine:1983), JHH-5 (TheTokyoJikeiUniversitySchoolofMedicine:1984), JHH-6 (TheTokyoJikeiUniversitySchoolofMedicine:1984), OHR (ShowaUniversity, schoolofmedicine:1985), KMCH-1 (KurumeUniversity, schoolofmedicine:1985), KMG-A (KurumeUniversity, schoolofmedicine:1985), JHH-7 (TheTokyoJikeiUniversitySchoolofMedicine:1986), JHC-1 (TheTokyoJikeiUniversitySchoolofMedicine:1986), KYN-1 (KurumeUniversity, schoolofmedicine:1986), KYN-2 (KurumeUniversity, schoolofmedicine:1987), HCC-T (KeioUniversity, schoolofmedicine:1986), HPT-NT/D3 (KyushuUniversity, facultyofmedicine:1986), Hep-tabata (MieUniversity, FacultyofMedicine:1986), HuCC-T1 (ToyamaMedicineandPharmaceuticalUniversity, facultyofmedicine:1987), HuH-28 (OkayamaUniversity, medicalschool:1987). above-mentioned relevant data can see human cell Vol.1, No.1, p.106-126, 1988.
Existing a large amount of research and utilization bioreactor culture liver cell carrys out manufacture of intraocular liver. Artificial technology makes progress in kidney, heart and lung transplantation. Such as, utilize artificial liver and other technologies to retain liver function for a long time and have been reported (AnandA.C., IndianJ.Gastroenterol., 2003,22Suppl2:S69-74; Uedaetal., ASAIOJ, 2003,49 (4): 401-6; Tillesetal., J.HepatobiliaryPancreat.Surg., 2002,9 (6): 686-96; Metab.BrainDis., 2005,20 (4): 327-35; AndParkandLee, J.BiosciBioeng., 2005,99 (4): 311-9). Such as, liver utility appliance also had relevant report (Luetal., TissueEng., 2005,11 (11-12): 1667-77; PlessandSauer, TransplantProc., 2005,37 (9): 3893-5; MillisandLosanoff, Nat.Clin.Pract.GastroenterolHepatol., 2005,2 (9): 398-405; AndGeorgeJ., J.Assoc.PhysiciansIndia, 2004,52:719-22). These liver utility appliance are often used in transplantation. Such as, artificial liver and liver utility appliance can allow explosive liver failure patient keep clear-headed and calm before surgery; Patient can be allowed stable after transplantation, particularly when not reacted by Reperfu-sion, it is possible to replace liver transplantation in some cases. In order to utilize biological artificial liver and liver assistor better, it is necessary to invent light small and exquisite bio-reactor. Therefore it is required that institute's culturing cell produces to be badly in need of and enough liver differential proteins in reactor. The shortage of Human normal hepatocyte makes liver cancer cell have very big potentiality in this application. Investigator has successfully utilized HepG2 cell to produce inhibitor of apoptosis protein Bcl-2(TeradaS. in this kind of device, J.Biosci.Bioeng., 2003,95 (2): 146-51).
Liver cancer is a kind of common tumour, and most liver cancer is caused by hepatitis virus, such as hepatitis B virus (HBV), hepatitis C virus (HCV); There is no effective therapy at present. HCV infection is a kind of main human infection's disease, there is no effective vaccine and prevents and treats. Existing part evidence demonstrates the importance that cellular immunization removes hepatitis virus, but antiviral response mechanism antibody-mediated in patient body is still not clear. Antibody in most patient body can neutralize virus antigen, but in serum, the content of neutralizing antibody and intensity are unknown, lack and support that the cell of virus infection or animal model limit the research to neutralizing antibody.
Although more existing clones deriving from people's liver cancer, but these existing clone majorities are that low differentiation and hepatocyte function are unsound. According to the situation that we understand, there is no the clone that a strain can support HBV and HCV infection at present, which also limits the research and development of antiviral and application. Utilize human primary hepatocyte to detect a small amount of HCV despite research group partly to copy, but also there is no clone (TagawaM.etal., J.GastroenterolHepatol, the 1995,10:523-527 that can support wild-type HCV massive duplication; ItoT.etal., J.Gen.Virol., 1996,77 (Pt.5): 1043-1054; ItoT.etal., Hepatology, 2001,34:566-572). 2005, Ji Ge research group successfully presented physiological period (HellerT.etal., Proc.Natl.Acad.Sci.USA, the 2005,102:2579-2583 of 2a type HCVJFH1 in clone; WakitaT.etal., Nat.Med., 2005,11 (7): 791-796; ZhongJ.etal., PNAs, 2005,102 (26): 9294-9; LindenbachB.D.etal., Science2005,309 (5734): 623-6).
At present, the animal model that uniquely can support HCV infection is chimpanzee (LanfordR.E.etal., Virology, 2002,293:1-9). The advantage of this kind of animal model is the real physiological cycle presenting virus, although (AlterM.J.etal., N.Eng.J.Med., 1999,341:556-562 different from the pathological manifestations infecting people; BassettS.E.etal., J.Virol., 1998,72:2589-2599), but this kind of model provides a large amount of explanation for the immune response of host after virus infection. The maximum restriction utilizing this kind of model is that gorilla source is few, and expensive. Tree is a kind of animalcule close to primates, is easy to adapt to laboratory environment. There are some researches show that tree also can infect various human viroid, comprise hepatitis virus (dieZ.C.etal., Virology, 1998,244:513-520).
The transgenic mouse of expression of HCV core protein has been used to research hepatic pathology and tumour generates (LemonS.M.etal., Trans.Am.Clin.Climatol.Assoc., 2000,111:146-156). Most transgenic animal do not show the toxicity to liver cell; a kind of models show is had to go out lymphocyte inflammation and hepatic necrosis (ZhaoX.etal.; J.Clin.Invest; 2002; 109:221-232), other two kinds of models show go out steatosis and liver cancer (MoriyaK.etal., J.Gen.Virol; 1997,78 (Pt.7): 1527-1531; MoriyaK.etal., Nat.Med., 1998,4:1065-1067). Research (ChisariF.V.etal., Science, the 1985,230:1157-1160 of the Immunological diseases genesis mechanism having utilized HBV transgene mouse model greatly open to be caused by HBV; ChisariF.V.etal., Hepatology, 1995,22:1316-1325). Transgenic mice is not yet widely used in HCV immunology, and the immunological tolerance of viral protein is become the problem utilizing mouse model to study.
Existing result of study shows above, obtains and has the hepatoma cell line of liver cancer similar functions and animal model is very favourable. The potential application of these clones is including, but not limited to screening and evaluation antitumor drug; Clone is cultivated HBV, HCV, the natural infection mode of simulated virus; Screening and evaluation antiviral; The metabolism kinetic energy of research liver cell.
Summary of the invention
The technical problem to be solved in the present invention overcomes the deficiencies in the prior art, a kind of high differentiation is provided, can cultivate, have the functional performance of liver in vitro, can be used for express cell pigment and liver cell differential protein human liver cancer cell system HLCZ02, the also application of this human liver cancer cell system of corresponding offer HLCZ02 in cell model, animal model, the preparation antiviral and antitumor drug of screening etc.
For solving the problems of the technologies described above, the technical scheme that the present invention proposes is a kind of human liver cancer cell system HLCZ02 that can be used for express cell pigment and liver cell differential protein, described human liver cancer cell system HLCZ02 for being preserved in China typical culture collection center (being called for short CCTCC) and preserving number is the human liver cancer cell system of CCTCCNO:C201310. The depositary institution of this human liver cancer cell system HLCZ02 is China typical culture collection center CCTCC, the address of depositary institution is positioned at Wuhan University of China, preservation date is on January 17th, 2013, and this human liver cancer cell system HLCZ02 is named as human liver cancer cell system HLCZ02.
In above-mentioned human liver cancer cell system HLCZ02, its karyomit(e) number is preferably 89～123.
In above-mentioned human liver cancer cell system HLCZ02, it is preferable that, described human liver cancer cell system HLCZ01 shows that after G-band chromosome karyotyping it exists karyomit(e) opposite sexization phenomenon, and has a marker chromosomes in cell caryogram.
Above-mentioned human liver cancer cell system HLCZ02, it belongs to the human liver cancer cell system of high differentiation on the one hand; On the other hand, this human liver cancer cell system HLCZ02 can express cell pigment and liver cell differential protein. The main phalangeal cell cytochrome p 450 of described cytopigment; Particularly preferred, described Cytochrome P450 comprises one or more in cytochrome C yP1A1, cytochrome C yP1B1, cytochrome C yP2C9. Described liver cell differential protein comprises at least one in albumin (ALB), alpha antitrypsin (AAT).
As a total technical conceive, the above-mentioned human liver cancer cell system HLCZ02 of the present invention can in growth medium vitro culture, and then predictive compound can be used to the carcinogenic of cell or teratogenesis Study. Preparation can be further used for by chemocarcinogenesis or teratogenesis Study, screen or evaluate various antitumor drug.
As a total technical conceive, due to human liver cancer cell system HLCZ02 and the human liver cell height homology of the present invention, therefore, the human liver cancer cell system HLCZ02 of the present invention can be applicable to the occasion needing to carry out a large amount of homologous cell experiment. Such as, apply as cell model in liver cell metabolism or drug metabolism study (such as assessing compound metabolic activation effect) in liver.
As a total technical conceive, the present invention also provides a kind of above-mentioned human liver cancer cell system HLCZ02 as the application of the cell model of support hepatites virus infections. By cultivating hepatitis virus in cell model, can the natural infection mode of simulated virus, the existence replication status of research hepatitis virus in liver cell. Based on this, the human liver cancer cell system HLCZ02 of the present invention also can apply in preparation, screening or evaluation antiviral.
As a total technical conceive, the present invention also provides a kind of above-mentioned human liver cancer cell system HLCZ02 implanting or proceed to after in animal body the application as animal model. Concrete, the human liver cancer cell system HLCZ02 of the present invention can be used for the animal model of non-human, is implanted by human liver cancer cell system HLCZ02 or proceeds to (such as mouse body is interior) in animal body, can be used for setting up animal (such as mouse) model supporting virus infection. The animal model utilizing the present inventor hepatoma cell line HLCZ02 to set up is conducive to carrying out hepatopathy Neo-Confucianism and hepatitis virus research.
As a total technical conceive, the above-mentioned human liver cancer cell system HLCZ02 of the present invention has important function and the characteristic of liver, ammonia can be converted into urea; And the function that ammonia metabolism is urea is become the favourable instrument manufacturing artificial liver or liver utility appliance, for clinical or scientific research. Therefore, the human liver cancer cell system HLCZ02 of the present invention also can apply in the preparation of artificial liver.
Except above-mentioned basic application mode, the above-mentioned human liver cancer cell system HLCZ02 of the present invention also can be used for human parasite's research etc.; Also can study its function entering foreign gene at transit cell.
Compared with prior art, it is an advantage of the current invention that: the present invention screens, obtains a kind of human liver cancer cell system HLCZ02 with initiative meaning, this human liver cancer cell system HLCZ02 not only belongs to the human liver cancer cell system of high differentiation, and can vitro culture in the medium;It has important function and the characteristic of liver, ammonia can be converted into urea, it is possible to express cell cytochrome p 450, CyP1A1, CyP1B1 and CyP2C9 albumen; Due to the property that the present inventor hepatoma cell line HLCZ02 has, make the human liver cancer cell system HLCZ02 of the present invention can be widely used in antiviral, the preparation of antitumor drug, research and development, screening and evaluation, can be used for the various clinical treatment biological products such as the biological artificial liver of preparation, the human liver cancer cell system HLCZ02 of the screening and culturing of the present invention is more conducive to research people's liver metabolism function and disease progression process than other animal liver cells.
A kind of human liver cancer cell system HLCZ02, its depositary institution is China typical culture collection center (being called for short CCTCC), and address is Wuhan University of China, and preserving number is CCTCCNO:C201310, and preservation date is on January 17th, 2013.
Accompanying drawing explanation
Fig. 1 detects Albumin(albumin ALB in three kinds of cells by RT-PCR in the embodiment of the present invention) and AATmRNA express comparison and detection result, its be under gel imaging instrument shooting the electrophorogram arrived.
Fig. 2 detects Albumin(albumin ALB in three kinds of cells by WesternBlot in the embodiment of the present invention) and the comparison and detection result of AAT protein expression.
Fig. 3 is the detected result that can cause apoptosis in the embodiment of the present invention with TRAIL process HLCZ02 cell.
Fig. 4 is human liver cancer cell system HLCZ02(2n=118 in the specific embodiment of the invention) chromosome karyotype analysis image results.
Fig. 5 is human liver cancer cell system HLCZ02(2n=89 in the specific embodiment of the invention) chromosome karyotype analysis image results.
Fig. 6 is human liver cancer cell system HLCZ02(2n=123 in the specific embodiment of the invention) chromosome karyotype analysis image results.
Embodiment
Below in conjunction with Figure of description and concrete preferred embodiment, the invention will be further described, but protection domain not thereby limiting the invention.
In following embodiment, if no special instructions, it is ordinary method. Experiment material used in following embodiment or biotechnological formulation, if no special instructions, be the conventional reagent being commercially available. Quantitative experiment in following examples, all arranges and repeats experiment for three times, results averaged.
Major experimental material used in embodiment and reagent:
(1) cell cultures: human liver cancer cell system HLCZ02(preservation), 100mm Tissue Culture Plate (Costar company), 60mm Tissue Culture Plate (Costar company), six porocyte culture plates (Costar company), DMEM substratum (Invitrogen company), penicillin and Streptomycin sulphate (Invitrogen company), glutamine (Invitrogen company), sheep blood serum (Invitrogen company), trypsin Invitrogen company), non-essential amino acid (Invitrogen company), 1 × PBS(Hyclone company);
(2) cell total rna extracts: TRIZOL(Invitrogen company), Virahol (Shanghai raw work), dehydrated alcohol (the raw work in Shanghai), the raw work in DEPC(Shanghai);
(3) ReverseTranscriptionPCR and regular-PCR: InvitrogenRT-PCR test kit (Invitrogen company), Taq enzyme (Taraka company);
(4) total protein of cell extracts: RIPA lysate (Thermo company), proteinase inhibitor (Merck company), determination of protein concentration reagent (Bio-Rad company);
(5) WesternBlot:PVDF film (Bio-Rad company), anti-ALB antibody (SANTACRUZ), anti-AAT antibody (SANTACRUZ), two anti-goatanti-mouseIgG(HRP) (Invitrogen company), WesternBlot chemical luminous substrate (Thermo company), WesternBlot exposure machine (Keda company).
Embodiment:
The present embodiment obtains the human liver cancer cell system HLCZ02 of a kind of the present invention, this human liver cancer cell system HLCZ02 has been preserved in China typical culture collection center, and preserving number is CCTCCNO:C201310, the address of depositary institution is positioned at Wuhan University of China, preservation date is on January 17th, 2013, and this human liver cancer cell system HLCZ02 is named as human liver cancer cell system HLCZ02.
The foundation of HLCZ02 clone and cultural method:
1. getting the fresh surgical sample of male sex's liver cancer patient excision, PBS washes once, and tissue block scalpel is cut into 1mm3Fritter be laid in the culture dish (100 millimeters) processed with collagen protein, add 8ml fresh culture, add Urogastron simultaneously, final concentration is 10ng/ml;
2. change fresh culture after cultivating 2 days, within later every 3 days, change fresh culture;
3. observe after culture dish grows cell clone, PBS washes a cell, drips on cell clone by 0.25% pancreatin, puts in incubator and takes out after about 1min, with moving liquid rifle suck fresh culture medium, the clone after digestion is rushed sucking-off, transferred to 12 orifice plate resume and cultivate;
4. band cell is transferred to after covering with in 12 orifice plates in six orifice plates, is extended in 10cm culture dish after covering with again;
5. by 1 × 107It is subcutaneous that individual cell is inoculated into NOD-SCID mouse, grows obviously swollen block, continue to cultivate according to human liver cancer tissue cultural method and obtain HLCZ02 cell after taking out swollen block after about 45 days.
Being detected by RT-PCR containing liver cell specific gene ALB and AAT in the human liver cancer cell system HLCZ02 of the present embodiment, detailed process is as follows:
The human liver cancer cell system HLCZ02 of human liver cancer cell system Huh7, the present invention and Chinese hamster ovary line CHO(is bought from U.S. ATCC) it is laid in six porocyte culture plates (300,000/hole), often kind of a cell spreads and accounts for holes; After cultivating 24h, three kinds of cells respectively get a hole, extract cell total rna with TRIZOL, then get 1 μ g total serum IgE reverse transcription and become cDNA, carry out PCR by template of 1 μ LcDNA, and amplification ALB and AAT gene, the result after amplification is as shown in Figure 1. As seen from Figure 1, human liver cancer cell system Huh7 and human liver cancer cell system HLCZ02 all can amplify liver cell specific gene ALB and AAT, but not hepatoma cell line CHO can not expand this two kinds of genes.
Being confirmed by WesternBlot to express liver cell differential protein ALB and AAT in the human liver cancer cell system HLCZ02 of the present embodiment, detailed process is as follows:
Get the three kinds of cells remaining three holes in above-mentioned six porocyte culture plates, use RIPABuffer lysing cell, cell pyrolysis liquid is placed in 15min on ice, then with centrifugal 15min(4 DEG C of 13000rpm), get supernatant and proceed in new EP pipe; Lowry method surveys protein concentration, get 50 μ g albumen respectively and carry out polyacrylamide gel electrophoresis (SDS-PAGE), then by the protein delivery on polyacrylamide gel on pvdf membrane, finally resist, with the two of Anti-ALB, Anti-AAT primary antibodie and band HRP mark, the expression detecting ALB and AAT in each protein sample, result is as shown in Figure 2, as seen from Figure 2, human liver cancer cell system Huh7 and HLCZ02 cell all there is ALB and AAT protein expression, but not hepatoma cell line CHO does not express this two kinds of albumen.
Can confirming by above experiment, the present invention screens the human liver cancer cell system HLCZ02 of acquisition not only containing liver cell specific gene ALB and AAT, and can express liver cell differential protein ALB and AAT.
The human liver cancer cell system HLCZ02 of above-mentioned the present invention has been carried out chromosome karyotype analysis by us, and this chromosome karyotype analysis is with reference to U.S. ATCC animal cell line chromosome karyotype analysis method. Censorship cell routine goes down to posterity, and respectively the cell (1000 metacinesis cells) of P2, P4, P7 generation is carried out chromosome sectioning, and common karyotype chromosome counts, and calculates karyomit(e) distribution frequency; G band dyeing observe, CCD imaging and carry out karyotyping with VideoTest-Karyo3.1 software.
In the human liver cancer cell system HLCZ02 of the present embodiment, chromosome karyotype analysis result display karyomit(e) number is generally 89～123.
In the human liver cancer cell system HLCZ02 of the present embodiment, human liver cancer cell system HLCZ02 shows that after G-band chromosome karyotyping it exists karyomit(e) opposite sexization phenomenon, and has a marker chromosomes in cell caryogram. We are for wherein the most typical three group chromosomes (2n=118,2n=89,2n=,123 tri-group chromosome) karyotyping, its analytical results as shown in Fig. 4～Fig. 6, by Fig. 4～Fig. 6 it may be seen that cell caryogram all has a marker chromosomes.
In addition, the above-mentioned human liver cancer cell system HLCZ02 of the present embodiment can in growth medium vitro culture, and then predictive compound can be used to the carcinogenic of cell or teratogenesis Study. Preparation can be further used for by chemocarcinogenesis or teratogenesis Study, screen or evaluate various antitumor drug. Fig. 3 processes HLCZ02 cell with TRAIL withering of cell can be caused to die, and then can be used for the screening of antitumor drug.
The above-mentioned human liver cancer cell system HLCZ02 of the present embodiment can implanting or proceed to after in animal body the application as animal model. Concrete, the human liver cancer cell system HLCZ02 of the present invention can be used for the animal model of non-human, is implanted by human liver cancer cell system HLCZ02 or proceeds to (such as mouse body is interior) in animal body, can be used for setting up animal (such as mouse) model supporting virus infection. We utilize the present inventor hepatoma cell line HLCZ02 to establish mouse model, to find to cause the generation of tumour in HLCZ02 cell infusion to immunodeficient mouse body, namely the human liver cancer cell system HLCZ02 of the present invention can be used for building anti-tumor virus animal model and applies.
Claims (3)
1. a human liver cancer cell system HLCZ02 is in the application screened or evaluate in antitumor drug, it is characterized in that, described human liver cancer cell system HLCZ02 is a kind of human liver cancer cell system HLCZ02 that can be used for express cell pigment and liver cell differential protein, described human liver cancer cell system HLCZ02 is preserved in China typical culture collection center and preserving number is the human liver cancer cell system of CCTCCNO:C201310, described liver cell differential protein comprises albumin, alpha antitrypsin, and described antitumor drug is TRAIL; The karyomit(e) number of described human liver cancer cell system HLCZ02 is 89～123; Described human liver cancer cell system HLCZ02 shows that after G-band chromosome karyotyping it exists karyomit(e) opposite sexization phenomenon, and has a marker chromosomes in cell caryogram.
2. a human liver cancer cell system HLCZ02 in liver cell metabolism or drug metabolism study as the application of cell model, described human liver cancer cell system HLCZ02 is a kind of human liver cancer cell system HLCZ02 that can be used for express cell pigment and liver cell differential protein, described human liver cancer cell system HLCZ02 is preserved in China typical culture collection center and preserving number is the human liver cancer cell system of CCTCCNO:C201310, and described liver cell differential protein comprises albumin, alpha antitrypsin.
3. a human liver cancer cell system HLCZ02 is supporting hepatites virus infections or preparation, screening or is evaluating in antiviral the application as cell model, described human liver cancer cell system HLCZ02 is a kind of human liver cancer cell system HLCZ02 that can be used for express cell pigment and liver cell differential protein, described human liver cancer cell system HLCZ02 is preserved in China typical culture collection center and preserving number is the human liver cancer cell system of CCTCCNO:C201310, and described liver cell differential protein comprises albumin, alpha antitrypsin.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310214644.6A CN103305467B (en) | 2013-05-31 | 2013-05-31 | A kind of human liver cancer cell system HLCZ02 and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310214644.6A CN103305467B (en) | 2013-05-31 | 2013-05-31 | A kind of human liver cancer cell system HLCZ02 and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103305467A CN103305467A (en) | 2013-09-18 |
| CN103305467B true CN103305467B (en) | 2016-06-15 |
Family
ID=49131189
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310214644.6A Active CN103305467B (en) | 2013-05-31 | 2013-05-31 | A kind of human liver cancer cell system HLCZ02 and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103305467B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110859840A (en) * | 2019-11-29 | 2020-03-06 | 湖南大学 | Application of nicotinic acid in preparing medicine for treating chronic hepatitis B |
| CN111440769B (en) * | 2020-02-25 | 2021-07-06 | 浙江大学 | Human hepatocyte hepatoma cell strain and application thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1338512A (en) * | 2001-09-20 | 2002-03-06 | 复旦大学附属中山医院 | Human liver cancer cell line |
| CN101693886A (en) * | 2009-08-10 | 2010-04-14 | 北京肿瘤医院 | Liver cancer cell line and application thereof |
| CN102465113A (en) * | 2010-11-12 | 2012-05-23 | 复旦大学附属中山医院 | Human hepatoma carcinoma cell line and application thereof |
| CN102690784A (en) * | 2011-03-22 | 2012-09-26 | 上海市肿瘤研究所 | Establishment and application of hepatocellular carcinoma cell line HCC-LY10 |
| CN102690785A (en) * | 2011-03-22 | 2012-09-26 | 上海市肿瘤研究所 | Establishment and application of hepatocellular carcinoma cell line HCC-LY5 |
-
2013
- 2013-05-31 CN CN201310214644.6A patent/CN103305467B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1338512A (en) * | 2001-09-20 | 2002-03-06 | 复旦大学附属中山医院 | Human liver cancer cell line |
| CN101693886A (en) * | 2009-08-10 | 2010-04-14 | 北京肿瘤医院 | Liver cancer cell line and application thereof |
| CN102465113A (en) * | 2010-11-12 | 2012-05-23 | 复旦大学附属中山医院 | Human hepatoma carcinoma cell line and application thereof |
| CN102690784A (en) * | 2011-03-22 | 2012-09-26 | 上海市肿瘤研究所 | Establishment and application of hepatocellular carcinoma cell line HCC-LY10 |
| CN102690785A (en) * | 2011-03-22 | 2012-09-26 | 上海市肿瘤研究所 | Establishment and application of hepatocellular carcinoma cell line HCC-LY5 |
Non-Patent Citations (5)
| Title |
|---|
| 人肝癌细胞系EH－H1的建立及其相关生物学特性;钱炎珍 等;《中国肿瘤生物治疗杂志》;20090228;第16卷(第1期);84-87 * |
| 人肝癌细胞系HC-108的建立及其生物学特征;陈葳 等;《细胞生物学杂志》;20020831;第24卷(第4期);249-252 * |
| 人肝癌细胞系HCC-9724的建立及其生物学特性;师长宏 等;《第四军医大学学报》;20000630;第21卷(第6期);709-712 * |
| 人肝癌细胞系HepG2生物学特性分析;陈富强 等;《内蒙古大学学报(自然科学版)》;20070531;第38卷(第5期);306-310 * |
| 稳定表达细胞色素P4501A1人肝癌细胞系的建立;庞莉萍 等;《中国医科大学学报》;20061031;第37卷(第5期);438-442 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103305467A (en) | 2013-09-18 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103275933B (en) | Human liver cancer cell line HLCZ01 and application thereof | |
| Kim et al. | Three-dimensional (3D) printing of mouse primary hepatocytes to generate 3D hepatic structure | |
| JP2019017393A5 (en) | ||
| Chen et al. | Human liver cancer organoids: Biological applications, current challenges, and prospects in hepatoma therapy | |
| Wijaya et al. | KLRG1+ natural killer cells exert a novel antifibrotic function in chronic hepatitis B | |
| CN103212071B (en) | Stem cell fusion model of carcinogenesis | |
| WO2015180636A1 (en) | Specific medium for long-term maintenance and proliferation subculture of human hepatocytes and culture method | |
| Kim et al. | Prolongation of liver-specific function for primary hepatocytes maintenance in 3D printed architectures | |
| CN109337858A (en) | The liver precursor like cell model in the primary hepatocyte source for hepatitis B virus infection, preparation method and application | |
| Bao et al. | Enhanced hepatic differentiation of rat bone marrow-derived mesenchymal stem cells in spheroidal aggregate culture on a decellularized liver scaffold | |
| CN101659941A (en) | Immortalized human liver cell line, preparation method and application thereof | |
| JP4711381B2 (en) | Novel human liver cancer cell line, method for obtaining this cell and use thereof | |
| CN108753686B (en) | Tissue engineering liver model, construction method and application thereof | |
| CN103305467B (en) | A kind of human liver cancer cell system HLCZ02 and application thereof | |
| CN103320385B (en) | Humanized's differentiated hepatoma cell strain HL1017 and construction process thereof | |
| CN111593026A (en) | Fluorescence-labeled human triple-negative breast cancer bone metastasis cell line | |
| Bhatia et al. | Introduction to animal tissue culture science | |
| CN104673743A (en) | Tissue block culture method for obtaining primary cell from animal tissue | |
| CN108823168A (en) | The method of culture medium and its primary liver cancer cell lines of source of people for establishing HBV infection | |
| CN103122334A (en) | Primary culture and purification method of intestinal epithelial cells of carassius auratus gibelio | |
| Yang et al. | A promising hepatocyte-like cell line, CCL-13, exhibits good liver function both in vitro and in an acute liver failure model | |
| CN104726400A (en) | Animal-source-free component culture method for differentiation from human pluripotent stem cells to germ cells | |
| Nagornykh et al. | Protocol for chronic hepatitis B virus infection mouse model development by patient-derived orthotopic xenografts | |
| CN101368170A (en) | In vitro together culture technique for hepatocyte and kupffer cell | |
| RU2272638C1 (en) | Biotransplant, method for its preparing and method for treatment of chronic hepatitis and liver cirrhosis (variants) |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |