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[肺腺癌细胞系XLA-07的建立与鉴定]

[Establishment and characterization of lung adenocarcinoma cell line XLA-07].

作者信息

Ma Li-ju, Wang Hong-zhi, Bian Li, Shao Wen-ping, Tang Rui-zhu, Wang Qin-qin, Jin Ke-wei

机构信息

First Affiliated Hospital of Kunming Medical College, Kunming 650032, China.

出版信息

Zhonghua Bing Li Xue Za Zhi. 2012 May;41(5):335-9. doi: 10.3760/cma.j.issn.0529-5807.2012.05.011.

Abstract

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 10(5) 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/c 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 G(0)/G(1), 15.1% in S and 8.0% in G(2)/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 cytometry. 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.

摘要

目的

建立并鉴定一株来自云南省宣威市一名女性患者的肺腺癌细胞系。

方法

获取肺腺癌手术标本,立即在含有10%胎牛血清、10⁵ U/L青霉素和100 mg/L链霉素的RPMI 1640培养基中培养。当细胞在培养中传代40次以上实现稳定增殖后,通过细胞形态学、核型分析、蛋白标志物表达[细胞角蛋白(CKs)、上皮膜抗原(EMA)和CD蛋白]、生长动力学、细胞周期阶段分布、有丝分裂指数、软琼脂集落形成、细胞侵袭以及在Balb/c裸鼠中的致瘤性等方面研究该细胞系的生物学特性。

结果

所建立的细胞系在相差显微镜下作为贴壁依赖的单层短梭形、多边形至上皮样细胞稳定培养超过80代,为期一年。透射电子显微镜下观察到微腺腔和紊乱的微丝。生长曲线呈“S”形,细胞群体每46.7小时倍增一次。有丝分裂指数为1.5%,集落形成率为8.3%。细胞周期分布为G(0)/G(1)期占76.9%,S期占15.1%,G(2)/M期占8.0%。该细胞系显示亚三倍体核型,众数为66条染色体,中位数为64条染色体。免疫组化显示细胞表达CK7、CK8、CK(泛)和EMA。流式细胞术显示细胞表面高水平表达CD13和CD59。细胞能够在体外穿透基质胶,但在裸鼠中未能形成稳定的异种移植瘤。

结论

成功从一名宣威肺癌患者建立了一株新的人肺腺癌细胞系,命名为XLA-07。

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