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Establishment and characterization of a canine chondrosarcoma cell line:Mango

动物疾病（英文） 2024年第3期4卷 186-197页

作者： Meilin Wang Xiao Wang Lixin He Hongbo Gao Wenxuan Li Huili Feng Qingyuan Zhao Wenwen Zhang Chengzong Li Bohan Zhang Changwei Qiu Department of Clinical Veterinary Medicine College ofVeterinary MedicineHuazhong Agricultural UniversityWuhan 430070China

In the global progress of bone tumor research,established stable and long-lasting transgenic chondrosarcoma(CSA)cell lines are rare,mainly of murine and human origin,while the establishment of canine CSA cell lines has yet to be *** study established a canine CSA cell line to facilitate the basic clinical study of canine *** five cases of canine osteolytic disease were collected,and more than 10 bone tumor samples from dogs with typical clinical signs were used for primary cell culture.A cell line with stable passaging for more than 100 generations and mouse tumorigenic ability was successfully *** to the clinical characteristics of the dog and the histopatho-logical results of the primary tumor,CSA was diagnosed,and the CSA cell line was designated ***-tochemical(IHC)results showed that the immunoreactivity of bone gamma-carboxyglutamate protein(BGLAP),secreted protein acidic and rich in cysteine(SPARC),alkaline phosphatase(ALPL),vimentin(VIM)and S100 were ***,the immunoreactivity of pan-cytokeratin(PCK),chromogranin A(CGA),and platelet endothelial cell adhesion molecule-1(CD31)was ***(IF)results showed that the protein expressions in the Mango cell line were consistent with the IHC identification of the primary *** Mango cell line's doubling time was 43.92 h,and the cell formation rate exceeded 20％.There were abnormal chromosome numbers,hetero staining with toluidine blue,and certain calcification *** could be passaged stably and continuously with-out changing the cell morphology and *** vivo,the cells were successfully injected into the nude mice model with a tumorigenic rate of 100％.The immunophenotype of the xenograft tumor was consistent with that of the primary ***,we effectively established a canine CSA cell *** a promising cell material,this cell line can be used to construct a tumor-bearing model conducive to the subsequent basic re

关键词： Canine chondrosarcoma cell line Establishment Characterization Tumorigenicity

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GFAP promoter directs lacZ expression specifically in a rat hepatic stellate cell line

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World Journal of Gastroenterology 2006年第5期12卷 723-730页

作者： Gunter Maubach Michelle Chin Chia Lim Institute of Bioengineering and Nanotechnology 31 Biopolis WayThe Nanos#04-01138669Singapore.

AIM： The GFAP was traditionally considered to be a biomarker for neural gila （mainly astrocytes and nonmyelinating Schwann cells）. Genetically, a 2.2-kb human GFAP promoter has been successfully used to target astrocytes in vitro and in vivo. More recently, GFAP was also established as one of the several makers for identifying hepatic stellate cells （HSC）. In this project, possible application of the same 2.2-kb human GFAP promoter for targeting HSC was investigated. METHODS： The GFAP-lacZ transgene was transfected into various cell lines （HSC, hepatocyte, and other nonHSC cell types）. The transgene expression specificity was determined by X-gal staining of the β-galactosidase activity. And the responsiveness of the transgene was tested with a typical pro-fibrotic cytokine TGF-β1. The expression of endogenous GFAP gene was assessed by real-time RT-PCR, providing a reference for the transgene expression. RESULTS： The results demonstrated for the first time that the 2.2 kb hGFAP promoter was not only capable of directing HSC-specific expression, but also responding to a known pro-fibrogenic cytokine TGF-β1 by upregulation in a doseand time-dependent manner, similar to the endogenous GFAP. CONCLUSION： In conclusion, these findings suggested novel utilities for using the GFAP promoter to specifically manipulate HSC for therapeutic purpose.

关键词： Promoter Regions (Genetics) Animals Base Sequence cell line DNA, Recombinant Gene Expression Glial Fibrillary Acidic Protein Hepatocytes Humans Lac Operon RNA, Messenger Rats Transfection Transforming Growth Factor beta Transforming Growth Factor beta1

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In vitro acute cytotoxicity of the neonicotinoid insecticide acetamiprid to FG cells, the gill cell line of flounder Paralichthy olivaceus

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Acta Oceanologica Sinica 2006年第5期25卷 135-140页

作者： SU Feng ZHANG Shicui YANG Ming LI Hongyan Department of Bioengineering ＆ Pharmaceutics Qingdao University of Science ＆ Technology Qingdao 266042 China Department of Marine Biology Ocean University of China Qingdao 266003 China

The in vitro acute cytotoxicity of the neonicotinoid insecticide acetamiprid to FG cell line, derived from the gill of flounder Paralichthys olizaceus, was tested by the three widely used endpoint bioassays-neutral red （NR） assay, tetrazolium （MTT） assay and cell protein assay. It was found that acetamiprid was increasingly toxic to FG cells at concentrations of 1 μg/cm^3 or above, and the inhibitory concentration 50% values for NR, MTF, and cell protein assays were 38.38, 36.27 and 32.03 μg/cm^3, respectively. This appeared to be the first report on the in vitro cytotoxicity of acetamiprid to non-mammalian vertebrate cells. Ultrastructural examination revealed that for the cells exposed to 60 μg/cm^3 acetamiprid for 48 h, their mitochondria were severely damaged with the cristae swelled up or disrupted, while their nuclei and rough endoplasmic reticlum （RER） appeared to be still normal. This suggests that mitochondria are possibly the primary target of acetamiprid.

关键词： acetamiprid cell line cytotoxicity flounder

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Establishment of a human hepatoma multidrug resistant cell line in vitro

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World Journal of Gastroenterology 2010年第18期16卷 2291-2297页

作者： Zhou, Yuan Ling, Xian-Long Li, Shi-Wei Li, Xin-Qiang Yan, Bin Third Mil Med Univ Xinqiao Hosp Dept Gastroenterol Chongqing 400037 Peoples R China

AIM:To establish a multidrug-resistant hepatoma cell line(SK-Hep-1),and to investigate its biological ***:A highly invasive SK-Hep-1 cell line of human hepatocellular carcinoma,also known as malignant hepatoma was incubated with a high concentration of cisplatin(CDDP) to establish a CDDP-resistant cell subline(SK-Hep-1/CDDP).The 50% inhibitory dose(IC50) values and the resistance indexes [(IC50 SK-Hep-1/CDDP)/(IC50 SK-Hep-1)] for other chemotherapeutic agents and the growth curve of cells were all evaluated using cell counting kit-8 *** distribution of the cell cycles were detected by flow *** of acquired multidrug resistance P-glycoprotein(MDR1,ABCB1) and multidrug resistance-associated protein 1(MRP1,ABCC1) was compared with that in parent cells by Western blotting and immunofluorescence combined with laser scanning confocal ***:The SK-Hep-1/CDDP cells(IC50 = 70.61 ± 1.06 μg/mL) was 13.76 times more resistant to CDDP than the SK-Hep-1 cells(IC50 = 5.13 ± 0.09 μg/mL),and CDDP-resistant cells also demonstrated cross-resistance to many anti-tumor agents such as doxorubicin,5-fluorouracil and *** morphologies were determined in both SK-Hep-1 and SK-Hep-1/CDDP *** cell cycle distribution of the SK-Hep-1/CDDP cell line exhibited a significantly increased percentage of cells in S(42.2% ± 2.65% vs 27.91% ± 2.16%,P cells,while the percentage of cells in the G0/G1 phase decreased(37.5% ± 5.05% vs 59.83% ± 3.28%,P cells exhibiting the MDR ***:Multiple drug resistance of multiple drugs in the human hepatoma cell line SK-Hep-1/CDDP was closely related to the overexpression of MDR1 and MRP1.

关键词： Hepatoma cell line Multidrug resistance In vitro Cisplatin

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Establishment and Characterization of a Fin cell line Derived from the Atlantic Salmon Salmo salar and Its Application to Fish Virology Study

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Journal of Ocean University of China 2022年第4期21卷 946-954页

作者： JIA Peng LIN Lirong XU La YI Meisheng JIA Kuntong School of Marine Sciences Sun Yat-sen UniversityGuangzhou 510000China Southern Marine Science and Engineering Guangdong Laboratory(Zhuhai) Zhuhai 519082China Zhuhai Key Laboratory of Marine Bioresources and Environment Sun Yat-sen UniversityGuangzhou 510000China Marine Biology Institute of Shandong Province Shandong Key Laboratory of Disease Control in MaricultureQingdao 266104China

Atlantic salmon(Salmo salar)is an important economic fish that is seriously threatened by various viruses.A cell line designated as ASF derived from the caudal fin tissue of Atlantic salmon was established and characterized in this *** cells grew well in Dulbecco’s modified Eagle’s medium(DMEM)containing 20%fetal bovine serum at 20℃.DNA sequencing and comparative analysis of the cytochrome B gene verified that the ASF cell line originated from Atlantic *** analysis indicated that the modal chromosome number of ASF cells was *** susceptibility test showed that ASF cells were susceptive to two important fish viruses,viral hemorrhagic septicemia virus(VHSV)and red-spotted grouper nervous necrosis virus(RGNNV).Viral replication in ASF cells was further confirmed by qRT-PCR and transmission electron ***,VHSV and RGNNV infections could induce the cellular responses in ASF cells,as indicated by the differential expression of cellular antiviral response-related genes,interferon-1 and *** conclusion,the newly established ASF cell line can be applied as an in vitro tool in fish virology and immunity studies.

关键词： Salmo salar cell line viral hemorrhagic septicemia virus nervous necrosis virus immune response

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Establishment,Characterization and Expression Pattern of a Spleen cell line,SMSP,Derived from Turbot(Scophthalmus maximus)

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中国海洋大学学报:英文版 2023年第5期22卷 1403-1411页

作者： XUE Ting LIU Yiping GE Xuefeng LI Chao School of Marine Science and Engineering Qingdao Agricultural UniversityQingdao 266109China

A turbot(Scophthalmus maximus)cell line named SMSP was obtained from the *** origin of the cells was identified by morphology,chromosome number and COI *** optimal basic medium,serum concentration and growth temperature of the cells were *** cell line is mainly composed of fibroblast-like *** of the SMSP cells contained 44 chromosomes,and the sequence of COI gene confirmed that the cells were originated from *** optimal culture conditions were 24℃,DMEM+10%*** cell line had high transfection efficiency for siRNA and *** stimulation with lipopolysaccharide(LPS)or poly(I:C),the expressions of immune-related genes such as TNF-β,IL-12s,IL-10 and IL-1βwere up-regulated significantly in the early stage(P<0.05).This study will provide a model for exploring immune mechanism of turbot against pathogen in vitro.

关键词： Scophthalmus maximus cell line spleen pathogen-associated molecular patterns immunity

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Establishment and characterization of an opisthorchiasis-associated cholangiocarcinoma cell line (KKU-100)

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World Journal of Gastroenterology 2005年第22期11卷 3392-3397页

作者： Banchob Sripa Saman Leungwattanawanit Takayuki Nitta Chaisiri Wongkham Vajarabhongsa Bhudhisawasdi Anucha Puapairoj Chongrak Sripa Masanao Miwa Department of Pathology Faculty of Medicine Khon Kaen University Khon Kaen 40002 Thailand Liver Fluke and Cholangiocarcinoma Research Center Khon Kaen University Khon Kaen 40002 Thailand Department of Pathology Faculty of Medicine Khon Kaen University Khon Kaen 40002 Thailand Institute of Basic Medical Science University of Tsukuba Ibaraki 305-8575 Tsukuba Japan Department of Biochemistry Faculty of Medicine Khon Kaen University Khon Kaen 40002 Thailand Liver Fluke and Cholangiocarcinoma Research Center Khon Kaen University Khon Kaen 40002 Thailand Department of Surgery Faculty of Medicine Khon Kaen University Khon Kaen 40002 Thailand Liver Fluke and Cholangiocarcinoma Research Center Khon Kaen University Khon Kaen 40002 Thailand Department of Pathology Faculty of Medicine Khon Kaen University Khon Kaen 40002 Thailand Puapairoj Liver Fluke and Cholangiocarcinoma Research Center Khon Kaen University Khon Kaen 40002 Thailand

AM: To establish and characterize a new cholangiocarcinoma cell line from a patient living in the Opisthorchis viverrini (O. viverrini) endemic area of Northeast Thailand. METHODS: Fresh liver biopsy and bile specimens were obtained from a 65-year-old Thai woman with cholangiocarcinoma of the ports hepatis. After digestion, the cells were cultured in Ham's F12 media. The established cell line was then characterized for growth kinetics, cell morphology, imm-unocytochemistry and cytogenetics. Tumorigenicity of the cell line was determined by heterotransplanting in nude mice. RESULTS: The primary tumor was a poorly differentiated tubular adenocarcinoma. Examination of the bile revealed malignant cells with O. viverrin eggs. The cholangiocarcinoma cell line KKU-100 was established 4 mo after the primary culture-population doubling time was 72 h. KKU-100 possesses compact and polygonal-shaped epithelial cells. Immunocytochemically, this cell line exhibited cytokeratin, EMA, CEA, and CA125, but not a-fetoprotein (AFP), CA19-9, desmin, c-met, or p53. Such protein expressions parallel those of the primary tumor. Cytogenetic analysis identified aneuploidy karyotypes with a modal chromosome number of 78 and marked chromosomal structural changes. Inoculation of KKU-100 cells into nude mice produced a transplantable, poorly differentiated aden-ocarcinoma, similar to the original tumor. CONCLUSION: KKU-100 is the first egg-proven, Opisthorchis- associated cholangiocarcinoma cell line, which should prove useful for further investigations of the tumor biology of this cancer.

关键词： Cholangiocarcinoma cell line Establishment Characterization Opisthorchiasis

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Establishment and characterization of a new cell line derived from human colorectal laterally spreading tumor

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World Journal of Gastroenterology 2008年第8期14卷 1204-1211页

作者： Xin-Ying Wang Zhou-Sheng Lai Chung-Man Yeung Ji-De Wang Wen Deng Hoi Yee Li Yu-Jing Han Hsiang-Fu Kung Bo Jiang Marie Chia-mi Lin Department of Gastroenterology Nanfang HospitalNanfang Medical University Tong He Guangzhou 510520Guangdong Province China Departmentof Chemistry the University of Hong Kong Pokfulam RoadHong Kong China Department of Anatomy the University of HongKong Pokfulam Road Hong Kong China Li Ka Shing Institute of Health Sciences The Centre for Emerging Infectious Diseases the Chinese Universityof Hong Kong Hong Kong China

AIM:To study the molecular mechanism of laterally spreading tumor (LST), a cell line [Laterally Spreading Tumor-Rectum 1 (LST-R1)] was derived and the characteristics of this cell line were investigated. METHODS:A new cell line (LST-R1) originated from laterally spreading tumor was established. Properties of the cell line were characterized using scanning and transmission electron microscopy, immunohistochemistry method, cytogenetic analysis and nude mice xenograft experiments. In vitro invasion assay, cDNA microarray and Western blotting were used to compare the difference between the LST-R1 and other colorectal cancer cell lines derived from prudent colon cancer. RESULTS:Our study demonstrated that both epithelial special antigen (ESA) and cytokeratin-20 (CK20) were expressed in LST-R1. The cells presented microvilli and tight junction with large nuclei. The karyotypic analysis showed hyperdiploid features with structural chromosome aberrations. The in vivo tumorigenicity was also demonstrated in nude mice xenograft experiments. The invasion assay suggested this cell line has a higher invasive ability. cDNA microarray and Western blotting show the loss of the expression of E-cadherin in LST-R1 ***:We established and characterized a colorectal cancer cell line, LST-R1 and LST-R1 has an obvious malignant tendency, which maybe partially attributed to the changes of the expression of some adhesion molecules, such as E-cadherin. It is also a versatile tool for exploring the original and progressive mechanisms of laterally spreading tumor and the early colon cancer genesis.

关键词： LST-R1 cell line Invasion E-cadherin

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Establishment and characterization of a cholangiocarcinoma cell line (RMCCA-1) from a Thai patient

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World Journal of Gastroenterology 2006年第40期12卷 6500-6506页

作者： Panthip Rattanasinganchan Kawin Leelawat Sa-ard Treepongkaruna Chintana Tocharoentanaphol Somboon Subwongchareon Tuangporn Suthiphongchai Rutaiwan Tohtong Department of Biochemistry Faculty of Science Mahidol University Bangkok 10400 Thailand Department of Biochemistry Faculty of Science Mahidol University Bangkok 10400 ThailandDepartment of Surgery Rajavithi Hospital Bangkok 10400 ThailandCancer Cytogenetic Unit Chulabhorn Cancer Center Chulabhorn Research Institute Patumthani 12000 Thailand Department of Surgery Rajavithi Hospital Bangkok 10400 Thailand Cancer Cytogenetic Unit Chulabhorn Cancer Center Chulabhorn Research Institute Patumthani 12000 Thailand

AIM: To establish and characterize a new cell line derived from peripheral cholangiocarcinoma of a Thai ***: The peripheral cholangiocarcinoma specimen surgically obtained from the patient was aseptically processed by washing and mincing before culturing in Ham’s F12 medium containing 10% fetal bovine serum. After 3 mo, when the cell line has become homogeneous and stabilized, several features were investigated, including growth characteristics, immunofluorescence staining for cytokeratins, expression of tumor markers, chromosomal analysis by G-banding and multicolour fluorescence in situ hybridization (mFISH), in vitro migration and invasion characteristics. RESULTS: The RMCCA-1 cell line has been established. These cells proliferated as a monolayer with a population doubling time of 48 h. Immunofluorescence staining showed positive staining for human cytokeratin 7 and 19 verifying the biliary epithelial origin. RMCCA-1 secreted carbohydrate antigen 19-9 (CA19-9), but insignificant levels of carcinoembryonic antigen (CEA) and α-fetoprotein (AFP). Chromosome analysis identified aneuploidy karyotypes with a modal chromosome number of 59. RMCCA-1 exhibited a low level of in vitro invasiveness, but a high degree of motility. The cell line exhibited a significant number of chromosomal aberrations as shown by mFISH and G-banding ***: A new cell line derived from peripheral cholangiocarcinoma of a Thai patient has been established. This cell line shows a low level of in vitro invasiveness, but a high degree of motility. It will serve as a valuable tool for further studies on tumor biology, molecular pathogenesis, metastatic mechanism and response to therapeutic drugs of cholangiocarcinoma.

关键词： Cholangiocarcinoma cell line Establishment mFISH Invasion Migration

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Contragestazol (DL111-IT) inhibits proliferation of human androgen-independent prostate cancer cell line PC3 in vitro and in vivo

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Asian Journal of Andrology 2005年第4期7卷 389-393,页

作者： Qiao-Jun He Bo Yang Yi-Jia Lou Rui-Ying Fang Department of Pharmacology College of Pharmaceutical Science Zhejiang University Hangzhou 310031 China

Aim： To evaluate the antiproliferative activity of contragestazol （DL111-IT） on the human prostate cancer cell line PC3 in vitro and in vivo and to elucidate its potential molecular mechanisms. Methods： The cell killing ability of DL111-IT was measured by the 3-（4,5-dimethylthia-zol,2-yl）-2,5-diphenyltetrazolium bromide （MTT） reagent assay method and the tumor xenograft model. The cell cycle was analyzed by flow cytometry and protein expression, including retinoblastoma （pRb）, cyclin-dependent kinase 4 （CDK4） and cyclin D 1, was detected by Western blotting. Results： DL111-IT exhibited high efficiency on cell growth inhibition of the human androgen-independent prostate cancer cell line PC3. The drug concentration that yielded 50 % cell inhibition （IC50 value） was 9.9 mg/mL. In the PC3 tumor xenograft study, DL111-IT （1.25 mg/kg-20.0 mg/kg） given once a day for 10 days significantly inhibited tumor growth, with the inhibition rate ranging from 21% to 50 %. Flow cytometric analysis indicated that DL111-IT could cause GI arrest in the PC3 cell line, but not apoptosis. DL111-IT enhanced pRb expression and down-regulated CDK4 and cyclin D 1 expression, suggesting that cell cycle regulation might contribute to the anticancer property of DL 111- IT. Conclusion： DL111-1T inhibits the proliferation of human androgen-independent prostate cancer cell line PC3 in vitro and in vivo by a cell cycle regulation pathway.

关键词： DL111-IT prostate cancer pRb cyclin-dependent kinase 4 cyclin D 1 PC3 cell line

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