宣威肺腺癌细胞系XLA-07的建立及特征

Establishment and characterization of lung adenocarcinoma cell line XLA-07

目的建立云南省宣威地区女性肺腺癌细胞系，为研究该地区女性肺癌发病机制提供体外实验模型。方法以手术切除的肺癌组织为标本，采用原代细胞培养法进行体外培养，建系后通过检测细胞形态、细胞生长曲线、群体倍增时间、细胞分裂指数、细胞周期和软琼脂克隆形成率、染色体核型、细胞角蛋白、CD分子、体外侵袭力和异种移植致瘤性等指标，分析鉴定所建细胞系的主要细胞生物学特征。结果新建立的XLA-07细胞生长稳定，可连续长期传代，形态呈梭形与多边形，异型性明显，电镜下可见微腺腔与排列紊乱的微丝，免疫细胞化学染色显示细胞角蛋白7、细胞角蛋白18、广谱角蛋白和上皮细胞膜抗原均呈阳性，细胞高表达CD59与CD13，细胞生长曲线呈“S”形，群体倍增时间约46．7h，细胞分裂指数为1．5％，软琼脂克隆形成率为8．3％，细胞周期G0／G1期占76．9％，G2／M期占8．0％，S期占15．1％，染色体主干系为亚三倍体核型，众数为66条，中位数64条，符合恶性腺上皮细胞特征。体外侵袭力检测发现细胞可穿过细胞外基质胶，但未能在裸鼠皮下形成稳定的移植瘤。无支原体污染。结论新建立了一个人肺腺癌细胞系，命名为XLA-07，该细胞系符合建系标准，是一个新建立的宣威女性肺腺癌细胞系。

Objective To establish and characterize a lung adenocarcinoma cell line from a female patient in Xuanwei, Yunnan province. Methods Surgical specimen of the lung adenocarcinoma was obtained and cultured immediately in RPMI 1640 medium with 10% fetal bovine serum and 105 U/L penicillin and 100 mg/L streptomycin. When stable proliferation of the cells was achieved after over 40 passages in culture, the biological features of the cell line were investigated by cell morphology, karyotyping, protein marker expression [ cytokeratins （ CKs ） , epithelial membrane antigen （ EMA ） and CD proteins ], growth kinetics, cell cycle phase distribution, mitotic index, colony formation in soft agar, cell invasion and tumorigenicity in Balb/e nude mice. Results The established cell line was stably cultured for over 80 passages during a one-year period as an anchorage-dependent monolayer of short spindle, polygonal to epithelioid cells under phase contrast microscope. Microglandular cavities and disordered microfilaments were observed under transmission electron microscope. The growth curve presented in an ＂S＂ shape with the cell population doubled every 46. 7 hours. The mitotic index was 1.5% and the colony formation rate was 8. 3%. The cell cycle distribution included 76. 9% in G0/G1 , 15.1% in S and 8.0% in G2/M. The cell line displayed a hypotriploid karyotype with a mode of 66 chromosomes and a median of 64 chromosomes. The cells expressed CK7, CK8, CK（Pan） and EMA by immunohistochemistry. A high level of cell surface expression of CD13 and CD59 was evident by flow eytometry. The cells were able to penetrate Matrigel in vitro but failed to form a stable xenograft in nude mice. Conclusion A new human lung adenocarcinoma cell line, designated as XLA-07, is successfully established from a Xuanwei lung cancer patient.