保留乳腺叶状瘤特征的乳腺叶状瘤细胞系的建立

Shishi He, Xiaoyun Xiao, R. Lei, Jiewen Chen, Hongyan Huang, Ailifeire Yilihamu, M. Guo, Cui Tan, Xun Li, Zilin Zhuang, Phei Er Saw, Yan Nie
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摘要

乳腺叶状瘤(PTs)是双相的,有上皮和间质成分。虽然PT的发病率很低(约占所有乳腺肿瘤的1%),但其临床结果难以预测,恶性PT往往进展迅速。目前没有有效的治疗方法,因此导致恶性PTs的死亡率很高。为了便于PT的研究,必须建立PT细胞系。本研究通过连续传代或细胞永生化方法建立了6株PT细胞系。我们通过体外功能测定、恶性PT标记检测和短串联重复序列鉴定来鉴定这些PT细胞系。用人乳头瘤病毒16 E6/E7转染良性PT细胞株SYSH-BPT-01和SYSH-BPT-02,用猴病毒40大T抗原转染恶性PT细胞株SYSH-MPT-01和SYSH-MPT-02。通过连续传代建立2株恶性PT细胞株SYSH-MPT-03和SYSH-MPT-04。所有恶性PT细胞株均表现出比良性PT细胞株更强的增殖、集落形成、迁移、侵袭和胶原收缩能力。此外,恶性PT标志物α-平滑肌肌动蛋白和成纤维细胞激活蛋白的表达水平和短串联重复序列鉴定表明,各PT细胞系与亲代原代细胞相同。我们成功建立了保留原代细胞特征的PT细胞系。这些细胞系可以作为研究乳腺PT功能的理想实验模型,从而为PT药物筛选和治疗靶点验证开辟新的可能性。
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Establishment of Breast Phyllodes Tumor Cell Lines Preserving the Features of Phyllodes Tumors
Breast phyllodes tumors (PTs) are biphasic, with epithelial and stromal components. Although the PT incidence is low (approximately 1% of all breast tumors), its clinical outcomes are unpredictable, and malignant PTs often progress rapidly. No effective treatment is currently available, thus resulting a high mortality rate from malignant PTs. PT cell lines must be established to facilitate the study of PTs. Herein, we established six PT cell lines through continuous passage or cell immortalization. We characterized these PT cell lines through in vitro functional assays, malignant PT marker detection and short tandem repeat identification. Benign PT cell lines (SYSH-BPT-01 and SYSH-BPT-02) were transfected with human papillomavirus 16 E6/E7, and two malignant PT cell lines (SYSH-MPT-01 and SYSH-MPT-02) were transfected with Simian virus 40 large T antigen. Two malignant PT cell lines (SYSH-MPT-03 and SYSH-MPT-04) were established through continuous passage. All malignant PT cell lines showed greater proliferation, colony formation, migration, invasion and collagen contraction ability than the benign PT cell lines. Moreover, the expression levels of malignant PT markers (α-smooth muscle actin and fibroblast activation protein) and short tandem repeat identification indicated that each PT cell line was identical to the parental primary cells. We successfully established PT cell lines that preserved the features of primary cells. These cell lines may serve as ideal experimental models for studying the function of breast PTs, thus opening new possibilities for PT drug screening and therapeutic target validation.
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