摘要
目的从1例中国女性肺癌患者的胸腔积液中分离建立原代低分化肺癌细胞系,并进行生物学特征分析。方法收集1例肺癌患者胸腔积液,取沉淀物,在含有10%胎牛血清、1%青霉素-链霉素溶液和1%非必需氨基酸的DMEM/F12培养液中培养。待细胞长满后,使用胰蛋白酶梯度消化法传代使肿瘤纯度达>95%。进一步分析已建立细胞系(命名为ZLC-001)的核型、短串联重复序列(short tandem repeats,STR)、细胞增殖、肿瘤标志物检测和成瘤性等生物学特征。肿瘤原代细胞三维立体培养法药敏检测技术(collagen gel droplet-embedded culture drug sensitivity test,CD-DST)用于评估ZLC-001对吉非替尼、舒尼替尼、索拉非尼、拉帕替尼和伊马替尼的药物敏感性。结果成功建立来自低分化肺癌患者(女性,44岁,pStageⅣ)的原代细胞系ZLC-001。ZLC-001细胞可贴壁生长,并可稳定传代>30代。生物学特征分析中核型分析表明染色体数目主要分布在60~69。免疫组织化学结果显示,细胞角蛋白(cytokeratin,CK)和乳腺癌相关蛋白1(breast cancer associated protein 1,BRAP1)为阳性,波形蛋白、Wnt-1和钙结合蛋白(calretinin,CR)个别阳性,Napsin A、MOC-31、甲状腺转录因子1(thyroid transcription factor 1,TTF1)和D2-40均为阴性。ZLC-001细胞在裸鼠中成瘤能力强。ZLC-001对吉非替尼、舒尼替尼、索拉非尼和伊马替尼耐药,但对拉帕替尼中度敏感。结论成功建立来自中国患者的新型低分化肺癌细胞系。这种功能齐全的细胞系为开展肺癌生物学和药物反应研究提供理想的模型与工具。
Objective To establish and characterize a poorly differentiated primary lung carcinoma cell line from a Chinese female lung cancer patient’s pleural effusions.Methods Pleural effusions were collected from a palliative paracentesis of a lung cancer patient.The sediment was cultured in DMEM-F12 medium(supplemented with 10%fetal bovine serum,1%penicillin-streptomycin solution and 1%non-essential amino acid).Differential trypsinization was used to get tumor purity above 95%when the cells were full.Established cell line was further analyzed for karyotype,short tandem repeats(STR),cell proliferation,tumor marker detection and tumorigenicity in nude.Collagen gel droplet-embedded culture drug sensitivity test(CD-DST)was used to evaluate the drug sensitivity of ZLC-001 to gefitinib,sunitinib,sorafernib,lapatinib and imatinib.Results A primary cell line from a poorly differentiated lung cancer patient(female,44 years old,pStageⅣ)was successfully established.ZLC-001 cells were adherent and maintained in culture for more than 30 passages.Karyotype examination showed that the number of chromosomes was mostly distributed between 60~69.The immunohistochemistry showed that cytokeratin(CK)and breast cancer associated protein 1(BRAP1)were positive,calretinin(CR),Wnt-1 and Vimentin were partially positive,and Napsin A,MOC-31,thyroid transcription factor 1(TTF1)and D2-40 were negative.Furthermore,ZLC-001 cells were highly tumorigenic in nude.ZLC-001 was resistant to gefitinib,sunitinib,sorafernib and imatinib,but medium sensitive to lapatinib.Conclusions A new poorly differentiated lung carcinoma cell line from a Chinese patient was successfully established.This well-characterized cell line provides a unique tool for future studies of investigating ethnic differences in lung cancer biology and drug response.
作者
黄旻然
金娇悦
吴映雪
徐晨阳
王金超
应莉莎
苏丹
Huang Minran;Jin Jiaoyue;Wu Yingxue;Xu Chenyang;Wang Jinchao;Ying Lisha;Su Dan(Department of Clinical Medicine,The Second Clinical Medical College of Zhejiang University of Traditional Chinese Medicine,Hangzhou 310053,China;Institute of Cancer and Basic Medicine(ICBM),Chinese Academy of Sciences,Department of Pathology,Cancer Hospital of the University of Chinese Academy of Sciences,Zhejiang Cancer Hospital,Hangzhou 310022,China;Institute of Cancer and Basic Medicine(ICBM),Chinese Academy of Sciences,Hangzhou 310022,China)
出处
《实用肿瘤杂志》
CAS
2020年第2期127-133,共7页
Journal of Practical Oncology
基金
国家自然科学基金(81972917)
浙江省部共建重大项目(WKJ-ZJ-1902-01)。
关键词
肺肿瘤
原代细胞系
吉非替尼耐药
lung neoplasms
primary cell lines
gefitinib resistant