CN101955911B - Chinese lung adenocarcinoma cell line with high metastases potentiality of bone, lever and adrenal gland - Google Patents
Chinese lung adenocarcinoma cell line with high metastases potentiality of bone, lever and adrenal gland Download PDFInfo
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Abstract
The invention belongs to the filed of microorganism animal cell lines and provides a Chinese lung adenocarcinoma cell line with high metastases potentiality of bone, lever and adrenal gland. According to the invention, hydrothorax of an adenocarcinoma first-diagnosis patient is used as primary culture of cells; the human adenocarcinoma cell has the preservation number of CGMCC No.3137 and classified name as human lung adenocarcinoma cell line CPA-Yang3, can grow adhered to the wall and has the tumorigenesis rate up to 100 percent; the human lung adenocarcinoma cell can grow quickly and metabolize vigorously; the expression levels of cancer genes EMS1, VEGF-C, IL-6, IL-8, SVIL and AR gene are higher than the expression level of SPC-A-1 cell; and the lung adenocarcinoma cancel cell has the biological characteristics of mainly transferring the bone and having high transfer potential of the lever and the adrenal gland, high growth speed of bone transfer cells and complete cell morphology. The invention can be used for providing reference data to early diagnosis for transferring the human lung adenocarcinoma bone, further establishing the relevant gene chip technology and assessing andexploring the curative effects of various medicaments and the like by comprehensively applying the gene chip, quantifying PCR (Polymerase Chain Reaction), western blot in real time and other technologies.
Description
Technical field
The invention belongs to biotechnology, technical field of microbe cell line, be specifically related to a kind of Chinese's lung adenocarcinoma cell line and uses thereof with bone, liver, suprarenal gland transfer high potential.
Background technology
Lung cancer is one of malignant disease of serious threat human survival and development, has become the No.1 killer in the tumor disease.2006, about 174470 people of lung cancer new case made a definite diagnosis in the U.S., accounted for 13% (male sex) and 12% (women) of whole tumours, and sickness rate is only second to skin carcinoma; Die from about 162460 people of lung cancer person, account for 31% (male sex) and 26% (women) of whole tumours, mortality ratio ranks first place
[1 ~ 2]The annual lung cancer new cases of Japan surpass 50000 people, mortality ratio nearly 90%
[3]Show that according to Shanghai City malignant tumour incidence survey in recent years lung cancer (21.56%) is in Shanghai male sex's malignant tumour sickness rate umber one, in the women, then is only second to mammary cancer
[4]The high for a long time major cause of lung cancer mortality is to be difficult to early diagnosis and its high transitivity, and many patient head have found many organ metastasis when examining
[3]Bone shifts and accounts for 30 ~ 70% of lung cancer patient, is only second to mammary cancer and prostate cancer
[5 ~ 7]Investigation shows that the patient above 1/3 can develop into osteolytic lesion, causes pain, pathologisch Bruch, spinal compression and hypercalcemia.Patient's quality of life is seriously descended, cause death at last, the course of disease generally is no more than 6 months
[3,8]Because the biological mechanism of bone metastases of lung cancer is still indeterminate so far, clinical shortage efficacious therapy method, therefore, studying the bone metastases of lung cancer pathogenesis and exploring effective diagnosis and treatment method is Chinese scholars striving direction for a long time.Although along with the development of basic subjects such as molecular biology, signal transduction path, protein function research, human research means to disease pathogenesis has had the development of advancing by leaps and bounds, because all objective restrictions, the research in this field does not have important breakthrough yet.
Research report is arranged, though host's microenvironment factor comprises that osteoclast and some have the stromatin of growth factor function etc. the transfer of tumour is had certain function
[3,9 ~ 10]Tumour cell self biological characteristics is undoubtedly the bone transfer and takes place more main and direct reason.
Because the operation taboo patient in late period is more common in the generation of bone metastases of lung cancer greatly, clinical samples is drawn materials limited, sluggish to a great extent bone metastases of lung cancer diagnosis and treatment progress of research.Therefore, if can set up effective people's bone metastases of lung cancer cell model, especially find and set up the bone metastasis model of parental generation human lung carcinoma cell; Can not only be more near human biological characteristics own; And the parental generation human lung adenocarcinoma cell model of the being built oncobiology characteristic that shifts of finder's adenocarcinoma of lung bone more effectively,, and then the functional gene that utilizes the institute's cell model of building research and development to be correlated with; Directly carry out intervention treatment research, its meliority is that clinical samples and other models are difficult to reach.
The theoretical basis of metastases animal model is that spontaneous transfer has relatively-stationary target tissue mostly in the tumour cell body.Find that after deliberation this characteristic is to be determined by a small amount of cell subset in the tumor tissues.Under certain conditions, such cell can form the metastases kitchen range at the particular target tissue by relatively-stationary pattern.Therefore, after many pickups kind immunodeficient animal was screened repeatedly with tumor cell line, this type of cell subset obtained effective enrichment, the progressive rising of the target tissue rate of transform.
No matter be the fundamental research of lung cancer; Comprise that pathology, immunology, genetics, pharmacogenomics, lung cancer functional genomics still are the clinic diagnosis practice of lung cancer; All require the investigator to walk the road of the research and development of oneself according to national conditions and compatriots' lung cancer morbidity characteristics.Cell is a Source of life, and tumour cell is the root of all evils.The lung carcinoma cell model of setting up the novel height transfer of China oneself will have significant scientific meaning and social value.The prior art relevant with the present invention has (to introduce in the preceding text):
Reference
[1].American?Cancer?Society.Cancer?facts?and?figures?2006.Atlanta，GA：American?CancerSociety；2006.pp.5~6
[2].Alex?G.Little?a，E.Greer?Gayb，Laurie?E.Gasparc，Andrew?K.Stewartd.National?surveyof?non-small?cell?lung?cancer?in?the?United?States：Epidemiology，pathology?and?patternsof?care.Lung?Cancer?2007，57(2)：253-260
[3].Saburo?Sene&Seiji?Yano.Molecular?pathogenesis?and?its?therapeutic?modalities?oflungcancer?metastasis?to?bone.Cancer?Metastasis?Rev.Springer?Science+Business?Media，LLC.2007
[4] urban district .2003 Shanghai City malignant tumour sickness rate. tumour, 2006,26 (7): 694.
[5].Coleman?RE.Metastatic?bone?disease：clinical?features，pathophysiology?andtreatment?strategies.Cancer?Treat?Rev?2001；27：165-176.
[6].G.David?Roodman.Mechanisms?of?Bone?Metastasis.NEJM.2004，350(4)：1655-1664
[7].Einat?Even-Sapir.Imaging?of?malignant?bone?involvement?by?morphologic，scintigraphic，and?hybric?modalities.J?Nucl?Med.2005；46(8)：1356~1367
[8].Katakami?N.Lung?cancer?with?bone?metastasis.Gan?To?Kagaku?Ryoho.2006；33(8)：1049-53.
[9].J.S.Nam，A.M.Suchar，M.J.Kang，et?al.Bone?Sialoprotein?Mediates?the?TumorGell-Targeted?Prometastatic?Activity?of?Transforming?Growth?Factor?beta?in?a?MouseModel?of?Breast?Cancer.Cancer?Res.2006；66(12)：6327-35.
[10].Hayashi?C，Rittling?S，Hayata?T，et?al.Serum?osteopontin，an?enhancer?of?tumormetastasis?to?bone，promotes?B16?melanoma?cell?migration.J?Cell?Biochem.2007；101(4)：979-86.
Summary of the invention
The purpose of this invention is to provide a kind of Chinese's lung adenocarcinoma cell line with bone, liver and suprarenal gland metastatic potential (is called for short: CPA-Yang3).Further purpose of the present invention provides the medical application of this lung cancer cell line.
The hydrothorax that the present invention adopts adenocarcinoma of lung head to examine patient (55 years old women) does that cell is former is commissioned to train fosterly, and this human lung adenocarcinoma cell confirms through experimentation on animals that this parental cell has had and only limits to the biological characteristics that bone shifts; This parental cell after the 5th generation in the mouse body, is induced after the nude inoculation cell suspension, and three Zhou Houneng detect 70% bone and shift; Dissect and find to have 50% hepatic metastases; 20% lung and suprarenal gland shift; After the amplification of bone transitional cell, bone video picture of nude mice nucleic and the film making of X line show that 80% mouse is after second circulation, and the video picture of nucleic bone detects the bone MET, and the mouse bone shifts and reaches more than 90% after the 3rd circulation.The present invention set up have bone, Chinese's lung adenocarcinoma cell line of liver and suprarenal gland metastatic potential (The Chinese Human Lung Adenocarcinoma cell with Highly Bone, Liverand Adrenal Gland Metastases, CPA-Yang3).This cell is in preservation on July 2 in 2009; Depositary institution: China Committee for Culture Collection of Microorganisms common micro-organisms center (CGMCC); No. 3, A, DaTun Road, Chaoyang District, BeiJing City, Datun Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica; 100101; Deposit number: CGMCC No.3137, the classification name: human lung adenocarcinoma cell line CPA-Yang3 (The Chinese HumanLung Adenocarcinoma cell with Highly Bone, Liver and Adrenal Gland Metastases, CPA-Yang3).
This cell strain has following biological characteristics:
1, adherent growth, the tumorigenesis rate is high
, gather fresh hydrothorax certainly and do and found all have the part cell to grow at the bottom of all cells culturing bottle in the 3rd day that cell cultivates; The 7th day, three bottles of cells that cover with at the bottom of the culturing bottle are used trysinization, get cell count and be about 4 * 10
6, process cell suspension after washing 2 times with saline water, concentration is 1 * 10
7/ mL.It is subcutaneous to be seeded in 2 immunodeficient mouse both sides respectively.Inoculate 60 days and still do not grow knurl.Reach just tumorigenesis and 100% long knurl of the 4th generation.Grow up to 3.5 * 4mm after one month
2With 4.7 * 28mm
2, 6.5 * 5.8mm
2, 7.2 * 5.7mm
2Tumorigenesis rate 100%.
2, to draw cell growth curve and show, growth is quick and metabolism is vigorous.
3, ESM1, VEGF-C, IL-6, IL-8, SVIL and AR gene expression dose,
Adopt the RT-PCR method these Chinese lung adenocarcinoma cell CPA-Yang3 and SPC-A-1 to be carried out the comparison of ESM1, VEGF-C, IL-6, IL-8, SVIL, AR and FN1 gene expression dose; The result shows that part oncogene ESM1, VEGF-C, IL-6, IL-8, SVIL, AR and the FN1 gene expression dose of these Chinese lung adenocarcinoma cell are apparently higher than the SPC-A-1 cell.
4, CPA-Yang3 is to be master's the cell strain with liver and suprarenal gland high metastatic potential with the bone transfer
Inoculate 70 * 10 respectively for after the parental cell after the 5th generation is induced in the mouse body every nude mice left ventricle
5/ 0.1mL cell suspension, three weeks can detect the transfer of 70% bone after create micropore pin hole living imaging method certainly; The when dissected naked eyes find to have 50% hepatic metastases; 20% lung and suprarenal gland shift; Get and make film for the back row nucleic bone video pictures of three week of nude mice left ventricle injection with the X line after the knee joint MET is cultivated the bone transitional cell and successfully increased again, 80% mouse still can be detected the bone MET and can not be detected by the X line behind second cycle by the video picture of nucleic bone.The bone MET that detects mainly concentrates on: mandibular bone, knee joint, humerus and backbone.The mouse left ventricle is inoculated 700,000 cells, 3 weeks of survival time.Has strong invasive ability.The mouse bone shifts and reaches more than 90% after the 3rd circulation.
5, bone transitional cell fast growth, cellular form is complete, has typical lotus knurl CPA-Yang3 bone and shifts mouse pathology figure.
The present invention has set up a Chinese's lung adenocarcinoma cell line with bone, liver and suprarenal gland metastatic potential.Through experiment confirm, Chinese Lu-csf-1 CPA-Yang3 is the very strong lung carcinoma cell of transfer ability.This cell through left ventricle injection only 700,000 cells, 2 week back mouse transfer appears and mouse seriously becomes thin, to such an extent as to the result that the two cage mouse of whole cycle are annihilated appearred in the present invention in experimentation.This experimental result has shown its favorable uniformity and repeatability; Bone transitional cell fast growth, cellular form is complete.Bone shifts and reaches more than 90% behind 3 in vivo-in vitro.It is a state's human lung carcinoma cell line that is worth research.Also be a good lung cancer metastasis cell model, shifting for systematic study lung cancer is multiple provides technology platform.
The present invention has set up high potential and has shifted the strain of human lung adenocarcinoma parental cell; Can carry out early diagnosis to the transfer of human lung adenocarcinoma bone reference data is provided; Also can further set up the genes involved chip technology; Technology such as integrated use gene chip, real-time quantitative PCR, Western blot (Western Blotting), assessment are explored various medicines and are comprised treatment curative effects such as chemistry, biology, immunity, tcm-, radiopharmaceuticals.For exploring diagnosis and treatment human body bone metastases of lung cancer technology platform is provided.
Simultaneously, further analyze the relevant functional genome of lung cancer metastasis and the genes involved of transfer, can deepen the understanding to the lung cancer metastasis molecule mechanism, the early diagnosis, the evaluation of curative effect that shift for bone provide a kind of effective molecular detecting method.
Description of drawings
Fig. 1 is the phase microscope figure of s-generation CPA-Yang3 cell (A * 100) and (B * 200).
Fig. 2 is the following plantation of the 4th generation cell skin 30 days.
Fig. 3 is the CPA-Yang3 cell growth curve.
Fig. 4 is the relatively expression level of CPA-Yang3 and SPC-A-1 portion gene ESM1, VEGF-c, IL-6, IL-8, SVIL, AR, FN1 of RT-PCR method.
Fig. 5 is that the mouse micropore pin hole bone video picture of CPA-Yang3 knee joint transitional cell left ventricle inoculation back is shown:
Wherein, A: left mandibular bone, left humerus, left knee joint and backbone shift, and the bone MET is done and grown again after cell is cultivated a week; B: bottom right jawbone, two humerus, cervical vertebra, backbone shift.
Fig. 6 is that CPA-Yang3 bone transitional cell inoculation back mouse bone shifts pathology, wherein, and A: knee joint (HE * 200); B: lumbar vertebrae (HE * 100).
Fig. 7 is CPA-Yang3 human lung adenocarcinoma bone transitional cell * 100.
Embodiment
Get hydrothorax that adenocarcinoma of lung head examines patient (55 years old women) do cell former be commissioned to train foster; Gather 50 milliliters of fresh light yellow hydrothorax; Every Tissue Culture Flask is inhaled 8 milliliters of hydrothorax and is added 2 milliliters of RPMI1640 nutrient solutions that do not contain Ox blood serum and shake up slightly after anti-with two, puts into culture temperature and is 37 ℃, CO2 concentration and be Germany of 5% and congratulate Li Shi BB16 type carbonic acid gas and incubate and support the case cultivation.Find to have approximately at the bottom of the Tissue Culture Flask 30% area to cover with cell when light microscopic is observed down after five days.Cell is adherent growth; Attached cell is Polygons mostly.Cell volume is bigger.Cultivate the incipient cell poor growth; Cellular form is stable, reaches for the 10th generation, and the cellular form and the speed of growth accelerate slightly.Proterties is constant behind the cryopreservation resuscitation.Observe CPA-Yang3 cell growing state differing under the inverted microscope: mostly attached cell is the Polygons that differs in size, and nuclear is big, kernel 2-3, visible division phase, and postmitotic cell still is adherent shape, and cell is obviously heterogeneous.Count 2 * 10 after the trysinization
5Cell adds a Tissue Culture Flask by conventional cell cultures.Under phase microscope, take cell growth picture.
This human lung adenocarcinoma cell confirms that through experimentation on animals this parental cell has had the biological characteristics that only limits to the bone transfer; Inoculate 70 * 10 respectively for after this parental cell after the 5th generation is induced in the mouse body every nude mice left ventricle
5/ 0.1mL cell suspension, three weeks can detect the transfer of 70% bone after create micropore pin hole living imaging method certainly; The when dissected naked eyes find to have 50% hepatic metastases; 20% lung and suprarenal gland shift; Get and make film for the back row nucleic bone video pictures of three week of nude mice left ventricle injection with the X line after the knee joint MET is cultivated the bone transitional cell and successfully increased again, 80% mouse can also be detected the bone MET and can not be detected by the X line behind second cycle by the video picture of nucleic bone.The bone MET that detects mainly concentrates on: mandibular bone, knee joint, humerus and backbone.The mouse left ventricle is inoculated 700,000 cells, 3 weeks of survival time.Has strong invasive ability.The mouse bone shifts and reaches more than 90% after the 3rd circulation.Set up have bone, Chinese's lung adenocarcinoma cell line of liver and suprarenal gland metastatic potential (The Chinese Human Lung Adenocarcinomacell with Highly Bone, Liver and Adrenal Gland Metastases, CPA-Yang3).
Embodiment 2
Above-mentioned human lung adenocarcinoma cell (CPA-Yang3) 70 * 10
4, inject the immunodeficient mouse blood system, weigh every other day after two weeks, rose in the 3rd week and carry out nucleic bone video picture detection weekly, the result confirms that this parental cell is that the high potential bone shifts lung carcinoma cell (Fig. 8).
Described substratum is for containing 10% foetal calf serum RPMI1640 or DMEM substratum, pH=7, and culture temperature is 37 ℃, 5%CO2.
Embodiment 3
RT-PCR detects demonstration: compare with the SPC-A-1 state human lung adenocarcinoma that prior art has been set up, part oncogene ESM1, VEGF-C, IL-6, IL-8, SVIL, AR and FN1 gene expression dose are apparently higher than the SPC-A-1 cell.
Claims (1)
1. Chinese's lung adenocarcinoma cell line with bone, liver and suprarenal gland metastatic potential; It is characterized in that said cell is the Chinese's lung adenocarcinoma cell line CPA-Yang3 with bone, liver and suprarenal gland metastatic potential; Deposit number: CGMCC No.3137, classification name: Chinese Lu-csf-1; Have following biological characteristics:
1) adherent growth, tumorigenesis rate height to 100%;
2) growth fast and metabolism vigorous;
3) oncogene ESM1, VEGF-C, IL-6, IL-8, SVIL and AR gene expression dose are higher than the SPC-A-1 cell;
4) be main with the bone transfer, have liver and suprarenal gland high metastatic potential;
5) bone transitional cell fast growth, cellular form is complete.
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| CN103131670B (en) * | 2011-11-26 | 2015-08-19 | 上海市胸科医院 | A kind of high potential brain metastes Chinese lung adenocarcinoma cell line |
| CN102424817B (en) * | 2011-12-31 | 2013-11-06 | 广州呼吸疾病研究所 | Cell strain from human lung adenocarcinoma and preparation method thereof |
| CN103756968B (en) * | 2013-11-20 | 2016-01-20 | 华东师范大学 | Spinal highly-metastatic human lung adenocarcinoma cell strain and construction process thereof and application |
| CN105567639A (en) * | 2014-10-13 | 2016-05-11 | 上海市肿瘤研究所 | Human lung adenocarcinoma cell LC-21 and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1712521A (en) * | 2004-12-15 | 2005-12-28 | 上海市胸科医院 | Lung cancer strain with high potential power of bone transference and its generation |
| CN1982441A (en) * | 2006-02-16 | 2007-06-20 | 上海市胸科医院 | Human lung adenocarcinoma self-transferring cell strain and its construction |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1712521A (en) * | 2004-12-15 | 2005-12-28 | 上海市胸科医院 | Lung cancer strain with high potential power of bone transference and its generation |
| CN1982441A (en) * | 2006-02-16 | 2007-06-20 | 上海市胸科医院 | Human lung adenocarcinoma self-transferring cell strain and its construction |
Non-Patent Citations (2)
| Title |
|---|
| Tang, GC等人.Lung adenocarcinoma metastasizing to cerebellopontine angle schwannoma (collision tumor).《ACTA NEUROCHIRURGICA 》.2007,第149卷(第1期),87-90. * |
| 杨顺芳等人.高转移性人肺腺癌细胞株SPC-A-1BM的建立及其特性分析.《肿瘤》.2006,第26卷(第12期),1059-1063. * |
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